Heterocyclic substituted hexahydropyrano[3,4-d][1,3]thiazin-2-amine compounds

ABSTRACT

Compounds, tautomers and pharmaceutically acceptable salts of the compounds are disclosed, wherein the compounds have the structure of Formula I, 
                         
as defined in the specification. Corresponding pharmaceutical compositions, methods of treatment, methods of synthesis, and intermediates are also disclosed.

This application is a Non-Provisional application, which claims the benefit of U.S. Provisional Patent Application No. 61/642,480 filed May 4, 2012.

FIELD OF THE INVENTION

The present inventions relate to small molecule inhibitors of 8-site amyloid precursor protein (APP) Cleaving Enzyme 1 (BACE1) and inhibitors of BACE2. In particular, this invention relates to inhibiting the production of A-beta peptides that can contribute to the formation of neurological deposits of amyloid protein, which may be applicable in the treatment of Alzheimer's Disease (AD) and other neurodegenerative and/or neurological disorders in mammals. In addition, this invention is related to the treatment of diabetes and obesity in mammals, including humans.

BACKGROUND OF THE INVENTION

Dementia results from a wide variety of distinctive pathological processes. The most common pathological processes causing dementia are AD, cerebral amyloid angiopathy (CM) and prion-mediated diseases (see, e.g., Haan et al., Clin. Neurol. Neurosurg. 1990, 92(4):305-310; Glenner et al., J. Neurol. Sci. 1989, 94:1-28). AD is a progressive, neurodegenerative disorder characterized by memory impairment and cognitive dysfunction. AD affects nearly half of all people past the age of 85, the most rapidly growing portion of the United States population. As such, the number of AD patients in the United States is expected to increase from about 4 million to about 14 million by 2050.

In addition, it has been determined that BACE1 knock-out mice had markedly enhanced clearance of axonal and myelin debris from degenerated fibers, accelerated axonal regeneration, and earlier reinnervation of neuromuscular junctions compared with littermate controls. These data suggest BACE1 inhibition as a therapeutic approach to accelerate regeneration and recovery after peripheral nerve damage. (See Farah, et al., J of Neurosci., 2011, 31(15): 5744-5754).

Insulin resistance and impaired glucose homoeostasis are important indicators of Type 2 diabetes and are early risk factors of AD. In particular, there is a higher risk of sporadic AD in patients with Type 2 diabetes and AD patients are more prone to Type 2 diabetes. It is believed that BACE1 levels may play a critical role in glucose and lipid homoeostasis in conditions of chronic nutrient excess. Consequently, the inhibition of BACE1 activity may also be important for the treatment of diabetes and obesity. Specifically, BACE1 inhibitors may be potentially useful for increasing insulin sensitivity in skeletal muscle and liver. (see Meakin et al., Biochem J. 2012, 441(1):285-96.)

Likewise, inhibition of BACE2 is proposed as a treatment of Type 2 diabetes with the potential to preserve and restore β-cell mass and stimulate insulin secretion in pre-diabetic and diabetic patients. (WO2011/020806). BACE2 is a β-cell enriched protease that regulates pancreatic β cell function and mass and is a close homologue of BACE1. Pharmacological inhibition of BACE2 increases β-cell mass and function, leading to the stabilization of Tmem27. (see Esterhazy et al., Cell Metabolism 2011, 14(3): 365-377). It is therefore an object of the present invention to provide for BACE2 inhibitors that are useful in the treatment and/or prevention of diseases associated with the inhibition of BACE2. (WO2011/020806).

Aminodihydrothiazine or thioamidine compounds are described in WO 2010/038686 as useful inhibitors of the β-secretase enzyme. The invention is directed to novel thioamidine compounds and their use in the treatment of neurodegenerative diseases, including AD, as well as the treatment of diabetes and obesity.

SUMMARY OF THE INVENTION

The present invention relates to:

(1) A compound represented by the Formula I:

wherein

R¹ is hydrogen or methyl, wherein said methyl is optionally substituted with one to three fluoro;

R² is a 5- to 6-membered heteroaryl, having one to three heteroatoms selected from N, O or S, wherein at least one of the heteroatoms is S or N and wherein said N is optionally substituted with R⁴; and wherein said heteroaryl moiety is independently substituted at each available carbon position with R^(3a), R^(3b), R^(3c) or R^(3d);

R^(3a), R^(3b), R^(3c) and R^(3d) are independently hydrogen, halogen, —OH, —CN, C₁₋₆alkyl, —OC₁₋₆alkyl; —(CR^(5a)R^(5b))_(m)—C₃₋₆cycloalkyl or —(CR^(5a)R^(5b))_(m)—C₃₋₆heterocycloalkyl; wherein said alkyl is optionally substituted with one to three fluoro and wherein said cycloalkyl and heterocycloalkyl moieties are optionally substituted with one to three fluoro, —CH₃, —CH₂F, —CHF₂ or —CF₃;

R⁴ is hydrogen, C₁₋₆alkyl, or —(CR^(5a)R^(5b))_(m)—C₃₋₆cycloalkyl; wherein said alkyl and cycloalkyl are optionally substituted with one to three fluoro, —OCH₃ or —OCF₃;

R^(5a) and R^(5b) are independently hydrogen, —CH₃, —CH₂F, —CHF₂, —CF₃ or —OCH₃; and

m is 0, 1 or 2;

or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer:

(2) A pharmaceutical composition comprising compounds of the invention and compounds of Formula I, or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer, or a solvate thereof, and a pharmaceutically acceptable vehicle, diluent or carrier;

(3) The pharmaceutical composition described herein for inhibiting production of amyloid-β protein and for inhibiting beta-site amyloid precursor protein cleaving enzyme 1 (BACE1);

(4) The pharmaceutical composition described herein for treating a neurodegenerative disease and, in particular, Alzheimer's Disease;

(5) The pharmaceutical composition described herein for inhibiting BACE1 and/or BACE2 activity for the therapeutic and/or prophylactic treatment of diseases and disorders characterized by elevated β-amyloid levels, including diabetes or type 2 diabetes;

(6) The pharmaceutical composition described herein for increasing insulin sensitivity in skeletal muscle and liver in a mammal, including humans;

(7) The pharmaceutical composition described herein for treating and/or preventing obesity.

(8) The compound or tautomer thereof or pharmaceutically acceptable salt of said compound or tautomer, or the solvate thereof, wherein the compound is selected from the compounds described in Table 7;

(9) Methods of inhibiting BACE2 enzyme activity, by administering a therapeutically effective amount of a thioamidine compound of any of the embodiments of Formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, to a mammal or a patient in need thereof.

(10) Methods for treating conditions or diseases of the central nervous system and neurological disorders in which the β-secretase enzyme is involved (such as migraine; epilepsy; Alzheimer's disease; Parkinson's disease; brain injury; stroke; cerebrovascular diseases (including cerebral arteriosclerosis, cerebral amyloid angiopathy, hereditary cerebral hemorrhage, and brain hypoxia-ischemia); cognitive disorders (including amnesia, senile dementia, HIV-associated dementia, Alzheimer's disease, Huntington's disease, Lewy body dementia, vascular dementia, drug-related dementia, tardive dyskinesia, myoclonus, dystonia, delirium, Pick's disease, Creutzfeldt-Jacob disease, HIV disease, Gilles de la Tourette's syndrome, epilepsy, muscular spasms and disorders associated with muscular spasticity or weakness including tremors, and mild cognitive impairment (“MCI”); mental deficiency (including spasticity, Down syndrome and fragile X syndrome); sleep disorders (including hypersomnia, circadian rhythm sleep disorder, insomnia, parasomnia, and sleep deprivation) and psychiatric disorders such as anxiety (including acute stress disorder, generalized anxiety disorder, social anxiety disorder, panic disorder, post-traumatic stress disorder, agoraphobia, and obsessive-compulsive disorder); factitious disorder (including acute hallucinatory mania); impulse control disorders (including compulsive gambling and intermittent explosive disorder); mood disorders (including bipolar I disorder, bipolar II disorder, mania, mixed affective state, major depression, chronic depression, seasonal depression, psychotic depression, seasonal depression, premenstrual syndrome (PMS) premenstrual dysphoric disorder (PDD), and postpartum depression); psychomotor disorder; psychotic disorders (including schizophrenia, schizoaffective disorder, schizophreniform, and delusional disorder); drug dependence (including narcotic dependence, alcoholism, amphetamine dependence, cocaine addiction, nicotine dependence, and drug withdrawal syndrome); eating disorders (including anorexia, bulimia, binge eating disorder, hyperphagia, obesity, compulsive eating disorders and pagophagia); sexual dysfunction disorders; urinary incontinence; neuronal damage disorders (including ocular damage, retinopathy or macular degeneration of the eye, tinnitus, hearing impairment and loss, and brain edema), nerve injury treatment (including accelerating regeneration and recovery after periphereal nerve damage) and pediatric psychiatric disorders (including attention deficit disorder, attention deficit/hyperactive disorder, conduct disorder, and autism) in a mammal, preferably a human, comprising administering to said mammal a therapeutically effective amount of a compound of Formula I or pharmaceutically acceptable salt thereof. The compounds of Formula I may also be useful for improving memory (both short-term and long-term) and learning ability. The text revision of the fourth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-IV-TR) (2000, American Psychiatric Association, Washington D.C.) provides a diagnostic tool for identifying many of the disorders described herein. The skilled artisan will recognize that there are alternative nomenclatures, nosologies, and classification systems for disorders described herein, including those as described in the DMS-IV-TR, and that terminology and classification systems evolve with medical scientific progress;

(11) Methods for treating a neurological disorder (such as migraine; epilepsy; Alzheimer's disease; Parkinson's disease; Niemann-Pick type C; brain injury; stroke; cerebrovascular disease; cognitive disorder; sleep disorder) or a psychiatric disorder (such as anxiety; factitious disorder; impulse control disorder; mood disorder; psychomotor disorder; psychotic disorder; drug dependence; eating disorder; and pediatric psychiatric disorder) in a mammal, preferably a human, comprising administering to said mammal a therapeutically effective amount of a compound of Formula I or pharmaceutically acceptable salt thereof;

(12) Methods for the treatment (e.g., delaying the progression or onset) of diabetes or diabetes-related disorders including Type 1 and Type 2 diabetes, impaired glucose tolerance, insulin resistance, hyperglycemia, and diabetic complications such as atherosclerosis, coronary heart disease, stroke, peripheral vascular disease, nephropathy, hypertension, neuropathy, and retinopathy;

(13) Methods for the treatment of obesity co-morbidities, such as metabolic syndrome. Metabolic syndrome includes diseases, conditions or disorders such as dyslipidemia, hypertension, insulin resistance, diabetes (e.g., Type 2 diabetes), coronary artery disease and heart failure. For more detailed information on metabolic syndrome, see, e.g., Zimmet, P. Z. et al., “The Metabolic Syndrome: Perhaps an Etiologic Mystery but Far From a Myth Where Does the International Diabetes Federation Stand?,” Diabetes & Endocrinology, 7(2), (2005); and Alberti, K. G. et al., “The Metabolic Syndrome—A New Worldwide Definition,” Lancet, 366, 1059-62 (2005);

(14) Methods for the treatment of nonalcoholic fatty liver disease (NAFLD) and hepatic insulin resistance;

(15) Combination therapies wherein the compounds of this invention may also be used in conjunction with other pharmaceutical agents for the treatment of the diseases, conditions and/or disorders described herein. Therefore, methods of treatment that include administering compounds of the present invention in combination with other pharmaceutical agents are also provided;

All patents, patent applications and references referred to herein are hereby incorporated by reference in their entirety.

Other features and advantages of this invention will be apparent from this specification and the appendent claims which describe the invention.

DEFINITIONS

The term “alkyl” refers to a linear or branched-chain saturated hydrocarbyl substituent (i.e., a substituent obtained from a hydrocarbon by removal of a hydrogen); in one embodiment from one to six carbon atoms; and in another embodiment, from one to four carbon atoms. Non-limiting examples of such substituents include methyl, ethyl, propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, sec-butyl and tert-butyl), pentyl, isoamyl, hexyl and the like.

The term “cycloalkyl” refers to a carbocyclic substituent obtained by removing a hydrogen from a saturated carbocyclic molecule and having three to six carbon atoms. In one embodiment, a cycloalkyl substituent has three carbon atoms. Non-limiting examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

In some instances, the number of carbon atoms in a hydrocarbyl substituent (i.e., alkyl, cycloalkyl, etc.) is indicated by the prefix “C_(x)-C_(y)-” or “C_(x-y)”, wherein x is the minimum and y is the maximum number of carbon atoms in the substituent. Thus, for example, “C₁-C₆-alkyl” or “C₁₋₆alkyl” refers to an alkyl substituent containing from 1 to 6 carbon atoms. Illustrating further, C₃-C₆-cycloalkyl refers to saturated cycloalkyl containing from 3 to 6 carbon ring atoms.

In some instances, the number of atoms in a cyclic substituent containing one or more heteroatoms (i.e., heteroaryl or heterocycloalkyl) is indicated by the prefix “x- to y-membered”, wherein x is the minimum and y is the maximum number of atoms forming the cyclic moiety of the substituent. Thus, for example, “5- to 6-membered heterocycloalkyl” refers to a heterocycloalkyl containing from 5 to 6 atoms, including one or more heteroatoms, in the cyclic moiety of the heterocycloalkyl. The heteroatoms for this invention are selected from N, O and S.

The term “hydroxy” or “hydroxyl” refers to —OH. When used in combination with another term(s), the prefix “hydroxy” indicates that the substituent to which the prefix is attached is substituted with one or more hydroxy substituents. Compounds bearing a carbon to which one or more hydroxy substituents include, for example, alcohols, enols and phenol.

The term “halo” or “halogen” refers to fluorine (which may be depicted as —F), chlorine (which may be depicted as —Cl), bromine (which may be depicted as —Br), or iodine (which may be depicted as —I).

The term “heterocycloalkyl” refers to a substituent obtained by removing a hydrogen from a saturated or partially saturated ring structure containing a total of 4 to 6 ring atoms, wherein at least one of the ring atoms is a heteroatom (i.e., oxygen, nitrogen, or sulfur), with the remaining ring atoms being independently selected from the group consisting of carbon, oxygen, nitrogen, and sulfur. In a group that has a heterocycloalkyl substituent, the ring atom of the heterocycloalkyl substituent that is bound to the group may be the heteroatom, or it may be a ring carbon atom. Similarly, if the heterocycloalkyl substituent is in turn substituted with a group or substituent, the group or substituent may be bound to a heteroatom, or it may be bound to a ring carbon atom.

The term “heteroaryl” refers to an aromatic ring structure containing from 5 to 6 ring atoms in which at least one of the ring atoms is a heteroatom (i.e., oxygen, nitrogen, or sulfur), with the remaining ring atoms being independently selected from the group consisting of carbon, oxygen, nitrogen, and sulfur. Examples of heteroaryl substituents include 6-membered ring substituents such as pyridyl, pyrazyl, pyrimidinyl, and pyridazinyl; and 5-membered ring substituents such as triazolyl, imidazolyl, furanyl, thiophenyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, 1,2,3-, 1,2,4-, 1,2,5-, or 1,3,4-oxadiazolyl and isothiazolyl. In a group that has a heteroaryl substituent, the ring atom of the heteroaryl substituent that is bound to the group may be one of the heteroatoms, or it may be a ring carbon atom. Similarly, if the heteroaryl substituent is in turn substituted with a group or substituent, the group or substituent may be bound to one of the heteroatoms, or it may be bound to a ring carbon atom. The term “heteroaryl” also includes pyridyl N-oxides and groups containing a pyridine N-oxide ring.

Examples of single-ring heteroaryls and heterocycloalkyls include furanyl, dihydrofuranyl, tetrahydrofuranyl, thiophenyl, dihydrothiophenyl, tetrahydrothiophenyl, pyrrolyl, isopyrrolyl, pyrrolinyl, pyrrolidinyl, imidazolyl, isoimidazolyl, imidazolinyl, imidazolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, triazolyl, tetrazolyl, dithiolyl, oxathiolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, thiazolinyl, isothiazolinyl, thiazolidinyl, isothiazolidinyl, thiaoxadiazolyl, oxathiazolyl, oxadiazolyl (including oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, or 1,3,4-oxadiazolyl), pyranyl (including 1,2-pyranyl or 1,4-pyranyl), dihydropyranyl, pyridinyl, piperidinyl, diazinyl (including pyridazinyl, pyrimidinyl, piperazinyl, triazinyl (including s-triazinyl, as-triazinyl and v-triazinyl), oxazinyl (including 2H-1,2-oxazinyl, 6H-1,3-oxazinyl, or 2H-1,4-oxazinyl), isoxazinyl (including o-isoxazinyl or p-isoxazinyl), oxazolidinyl, isoxazolidinyl, oxathiazinyl (including 1,2,5-oxathiazinyl or 1,2,6-oxathiazinyl), oxadiazinyl (including 2H-1,2,4-oxadiazinyl or 2H-1,2,5-oxadiazinyl), morpholinyl.

Additional examples of heteroaryls and heterocycloalkyls include: 3-1H-benzimidazol-2-one, 2-tetrahydrofuranyl, 3-tetrahydrofuranyl, 2-tetrahydropyranyl, 3-tetrahydropyranyl, 4-tetrahydropyranyl, [1,3]-dioxalanyl, [1,3]-dithiolanyl, [1,3]-dioxanyl, 2-tetrahydrothiophenyl, 3-tetrahydrothiophenyl, 2-morpholinyl, 3-morpholinyl, 4-morpholinyl, 2-thiomorpholinyl, 3-thiomorpholinyl, 4-thiomorpholinyl, 1-pyrrolidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 1-piperazinyl, 2-piperazinyl, 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-piperidinyl, 4-thiazolidinyl, pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidino, morpholino, thiomorpholino, thioxanyl, piperazinyl, azetidinyl, oxetanyl, thietanyl, oxazepinyl, 1,2,3,6-tetrahydropyridinyl, 2-pyrrolinyl, 3-pyrrolinyl, 2H-pyranyl, 4H-pyranyl, dioxanyl, 1,3-dioxolanyl, pyrazolinyl, dithianyl, dithiolanyl, dihydropyranyl, dihydrothienyl, dihydrofuranyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, 3-azabicyclo[3.1.0]hexanyl, 3-azabicyclo[4.1.0]heptanyl, pyridinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, pyridazinyl, triazinyl, oxadiazolyl, thiadiazolyl, furazanyl. The foregoing groups, as derived from the groups listed above, may be C-attached or N-attached where such is possible. For instance, a group derived from pyrrole may be pyrrol-1-yl (N-attached) or pyrrol-3-yl (C-attached). Further, a group derived from imidazole may be imidazol-1-yl (N-attached) or imidazol-2-yl (C-attached).

If substituents are described as being “independently selected” from a group, each instance of a substituent is selected independent of the other. Each substituent therefore may be identical to or different from the other substituent(s).

As used herein, the term “Formula I” may be hereinafter referred to as a “compound(s) of the invention.” Such terms are also defined to include all forms of the compound of Formula I, including hydrates, solvates, isomers, crystalline and non-crystalline forms, isomorphs, polymorphs, and metabolites thereof. For example, the compounds of the invention, or pharmaceutically acceptable salts thereof, may exist in unsolvated and solvated forms. When the solvent or water is tightly bound, the complex will have a well-defined stoichiometry independent of humidity. When, however, the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water/solvent content will be dependent on humidity and drying conditions. In such cases, non-stoichiometry will be the norm.

The compounds of the invention may exist as clathrates or other complexes. Included within the scope of the invention are complexes such as clathrates, drug-host inclusion complexes wherein, in contrast to the aforementioned solvates, the drug and host are present in stoichiometric or non-stoichiometric amounts. Also included are complexes of the compounds of the invention containing two or more organic and/or inorganic components which may be in stoichiometric or non-stoichiometric amounts. The resulting complexes may be ionized, partially ionized, or non-ionized. For a review of such complexes, see J. Pharm. Sci., 64 (8), 1269-1288 by Haleblian (August 1975).

The compounds of the invention have asymmetric carbon atoms. The carbon-carbon bonds of the compounds of the invention may be depicted herein using a solid line (-), a solid wedge (

), or a dotted wedge (

). The use of a solid line to depict bonds to asymmetric carbon atoms is meant to indicate that all possible stereoisomers (e.g., specific enantiomers, racemic mixtures, etc.) at that carbon atom are included. The use of either a solid or dotted wedge to depict bonds to asymmetric carbon atoms is meant to indicate that only the stereoisomer shown is meant to be included. It is possible that compounds of Formula I may contain more than one asymmetric carbon atom. In those compounds, the use of a solid line to depict bonds to asymmetric carbon atoms is meant to indicate that all possible stereoisomers are meant to be included. For example, unless stated otherwise, it is intended that the compounds of Formula I can exist as enantiomers and diastereomers or as racemates and mixtures thereof. The use of a solid line to depict bonds to one or more asymmetric carbon atoms in a compound of Formula I and the use of a solid or dotted wedge to depict bonds to other asymmetric carbon atoms in the same compound is meant to indicate that a mixture of diastereomers is present.

Stereoisomers of Formula I include cis and trans isomers, optical isomers such as R and S enantiomers, diastereomers, geometric isomers, rotational isomers, conformational isomers, and tautomers of the compounds of the invention, including compounds exhibiting more than one type of isomerism; and mixtures thereof (such as racemates and diastereomeric pairs). Also included are acid addition or base addition salts wherein the counterion is optically active, for example, D-lactate or L-lysine, or racemic, for example, DL-tartrate or DL-arginine.

When any racemate crystallizes, crystals of two different types are possible. The first type is the racemic compound (true racemate) referred to above wherein one homogeneous form of crystal is produced containing both enantiomers in equimolar amounts. The second type is the racemic mixture or conglomerate wherein two forms of crystal are produced in equimolar amounts each comprising a single enantiomer.

The compounds of Formula I may exhibit the phenomenon of tautomerism and are regarded as compounds of the invention. For example, the compounds of Formula I may exist in several tautomeric forms, including the 2-aminodihydrothiazine form, Ia, and the 2-imino-tetrahydrothiazine form, Ib. All such tautomeric forms, and mixtures thereof, are included within the scope of compounds of Formula I. Tautomers exist as mixtures of a tautomeric set in solution. In solid form, usually one tautomer predominates. Even though one tautomer may be described, the present invention includes all tautomers of the compounds of Formula I and salts thereof. Examples of tautomers are described by the compounds of Formula Ia and Ib and, collectively and generically are referred to as compounds of Formula I.

The compounds of this invention may be used in the form of salts derived from inorganic or organic acids. Depending on the particular compound, a salt of the compound may be advantageous due to one or more of the salt's physical properties, such as enhanced pharmaceutical stability in differing temperatures and humidities, or a desirable solubility in water or oil. In some instances, a salt of a compound also may be used as an aid in the isolation, purification, and/or resolution of the compound.

Where a salt is intended to be administered to a patient (as opposed to, for example, being used in an in vitro context), the salt preferably is pharmaceutically acceptable. The term “pharmaceutically acceptable salt” refers to a salt prepared by combining a compound of Formula I with an acid whose anion, or a base whose cation, is generally considered suitable for human consumption. Pharmaceutically acceptable salts are particularly useful as products of the methods of the present invention because of their greater aqueous solubility relative to the parent compound. For use in medicine, the salts of the compounds of this invention are non-toxic “pharmaceutically acceptable salts.” Salts encompassed within the term “pharmaceutically acceptable salts” refer to non-toxic salts of the compounds of this invention which are generally prepared by reacting the free base with a suitable organic or inorganic acid.

Suitable pharmaceutically acceptable acid addition salts of the compounds of the present invention when possible include those derived from inorganic acids, such as hydrochloric, hydrobromic, hydrofluoric, boric, fluoroboric, phosphoric, metaphosphoric, nitric, carbonic, sulfonic, and sulfuric acids, and organic acids such as acetic, benzenesulfonic, benzoic, citric, ethanesulfonic, fumaric, gluconic, glycolic, isothionic, lactic, lactobionic, maleic, malic, methanesulfonic, trifluoromethanesulfonic, succinic, toluenesulfonic, tartaric, and trifluoroacetic acids. Suitable organic acids generally include, for example, aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids.

Specific examples of suitable organic acids include acetate, trifluoroacetate, formate, propionate, succinate, glycolate, gluconate, digluconate, lactate, malate, tartaric acid, citrate, ascorbate, glucuronate, maleate, fumarate, pyruvate, aspartate, glutamate, benzoate, anthranilate, stearate, salicylate, p-hydroxybenzoate, phenylacetate, mandelate, embonate (pamoate), methanesulfonate, ethanesulfonate, benzenesulfonate, pantothenate, toluenesulfonate, 2-hydroxyethanesulfonate, sufanilate, cyclohexylaminosulfonate, algenic acid, β-hydroxybutyric acid, galactarate, galacturonate, adipate, alginate, butyrate, camphorate, camphorsulfonate, cyclopentanepropionate, dodecylsulfate, glycoheptanoate, glycerophosphate, heptanoate, hexanoate, nicotinate, 2-naphthalesulfonate, oxalate, palmoate, pectinate, 3-phenylpropionate, picrate, pivalate, thiocyanate, and undecanoate.

Furthermore, where the compounds of the invention carry an acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts, i.e., sodium or potassium salts; alkaline earth metal salts, e.g., calcium or magnesium salts; and salts formed with suitable organic ligands, e.g., quaternary ammonium salts. In another embodiment, base salts are formed from bases which form non-toxic salts, including aluminum, arginine, benzathine, choline, diethylamine, diolamine, glycine, lysine, meglumine, olamine, tromethamine and zinc salts.

Organic salts may be made from secondary, tertiary or quaternary amine salts, such as tromethamine, diethylamine, N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. Basic nitrogen-containing groups may be quaternized with agents such as lower alkyl (C₁-C₆) halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides), dialkyl sulfates (i.e., dimethyl, diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g., decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides), arylalkyl halides (e.g., benzyl and phenethyl bromides), and others.

In one embodiment, hemisalts of acids and bases may also be formed, for example, hemisulfate and hemicalcium salts.

Also within the scope of the present invention are so-called “prodrugs” of the compound of the invention. Thus, certain derivatives of the compound of the invention which may have little or no pharmacological activity themselves can, when administered into or onto the body, be converted into the compound of the invention having the desired activity, for example, by hydrolytic cleavage. Such derivatives are referred to as “prodrugs.” Further information on the use of prodrugs may be found in “Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella) and “Bioreversible Carriers in Drug Design,” Pergamon Press, 1987 (ed. E. B. Roche, American Pharmaceutical Association). Prodrugs in accordance with the invention can, for example, be produced by replacing appropriate functionalities present in the compounds of any of Formula I with certain moieties known to those skilled in the art as “pro-moieties” as described, for example, in “Design of Prodrugs” by H. Bundgaard (Elsevier, 1985).

The present invention also includes isotopically labeled compounds, which are identical to those recited in Formula I, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine and chlorine, such as ²H, ³H, ¹³C, ¹¹C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³²P, ³⁵S, ¹⁸F, and ³⁶Cl, respectively. Compounds of the present invention, prodrugs thereof, and pharmaceutically acceptable salts of said compounds or of said prodrugs which contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of this invention. Certain isotopically labeled compounds of the present invention, for example those into which radioactive isotopes such as ³H and ¹⁴C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., ³H, and carbon-14, i.e., ¹⁴C, isotopes are particularly preferred for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium, i.e., ²H, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically labeled compounds of Formula I of this invention and prodrugs thereof can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples and Preparations below, by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.

As used herein, “eating disorders” refer to illnesses in which the patient suffers disturbances in his/her eating behaviors and related thoughts and emotions. Representative examples of obesity-related eating disorders include overeating, bulimia, binge-eating disorder, compulsive dieting, nocturnal sleep-related eating disorder, pica, Prader-Willi syndrome, and night-eating syndrome.

DETAILED DESCRIPTION OF THE INVENTION

Typically, a compound of the invention is administered in an amount effective to treat a condition as described herein. The compounds of the invention are administered by any suitable route in the form of a pharmaceutical composition adapted to such a route, and in a dose effective for the treatment intended. Therapeutically effective doses of the compounds required to treat the progress of the medical condition are readily ascertained by one of ordinary skill in the art using preclinical and clinical approaches familiar to the medicinal arts.

The term “treating”, as used herein, unless otherwise indicated, means reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. The term “treatment”, as used herein, unless otherwise indicated, refers to the act of treating as “treating” is defined immediately above. The term “treating” also includes adjuvant and neo-adjuvant treatment of a subject.

The compounds of the invention may be administered orally. Oral administration may involve swallowing, so that the compound enters the gastrointestinal tract, or buccal or sublingual administration may be employed, by which the compound enters the blood stream directly from the mouth.

In another embodiment, the compounds of the invention may also be administered directly into the blood stream, into muscle, or into an internal organ. Suitable means for parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular and subcutaneous. Suitable devices for parenteral administration include needle (including microneedle) injectors, needle-free injectors and infusion techniques.

In another embodiment, the compounds of the invention may also be administered topically to the skin or mucosa, that is, dermally or transdermally. In another embodiment, the compounds of the invention can also be administered intranasally or by inhalation. In another embodiment, the compounds of the invention may be administered rectally or vaginally. In another embodiment, the compounds of the invention may also be administered directly to the eye or ear.

The dosage regimen for the compounds and/or compositions containing the compounds is based on a variety of factors, including the type, age, weight, sex and medical condition of the patient; the severity of the condition; the route of administration; and the activity of the particular compound employed. Thus the dosage regimen may vary widely. Dosage levels of the order from about 0.01 mg to about 100 mg per kilogram of body weight per day are useful in the treatment of the above-indicated conditions. In one embodiment, the total daily dose of a compound of the invention (administered in single or divided doses) is typically from about 0.01 to about 100 mg/kg. In another embodiment, total daily dose of the compound of the invention is from about 0.1 to about 50 mg/kg, and in another embodiment, from about 0.5 to about 30 mg/kg (i.e., mg compound of the invention per kg body weight). In one embodiment, dosing is from 0.01 to 10 mg/kg/day. In another embodiment, dosing is from 0.1 to 1.0 mg/kg/day. Dosage unit compositions may contain such amounts or submultiples thereof to make up the daily dose. In many instances, the administration of the compound will be repeated a plurality of times in a day (typically no greater than 4 times). Multiple doses per day typically may be used to increase the total daily dose, if desired.

For oral administration, the compositions may be provided in the form of tablets containing from about 0.01 mg to about 500 mg of the active ingredient, or in another embodiment, from about 1 mg to about 100 mg of active ingredient. Intravenously, doses may range from about 0.1 to about 10 mg/kg/minute during a constant rate infusion.

Suitable subjects according to the present invention include mammalian subjects. Mammals according to the present invention include, but are not limited to, canine, feline, bovine, caprine, equine, ovine, porcine, rodents, lagomorphs, primates, and the like, and encompass mammals in utero. In one embodiment, humans are suitable subjects. Human subjects may be of either gender and at any stage of development.

In another embodiment, the invention comprises the use of one or more compounds of the invention for the preparation of a medicament for the treatment of the conditions recited herein.

For the treatment of the conditions referred to above, the compound of the invention can be administered as compound per se. Alternatively, pharmaceutically acceptable salts are suitable for medical applications because of their greater aqueous solubility relative to the parent compound.

In another embodiment, the present invention comprises pharmaceutical compositions. Such pharmaceutical compositions comprise a compound of the invention presented with a pharmaceutically acceptable carrier. The carrier can be a solid, a liquid, or both, and may be formulated with the compound as a unit-dose composition, for example, a tablet, which can contain from 0.05% to 95% by weight of the active compounds. A compound of the invention may be coupled with suitable polymers as targetable drug carriers. Other pharmacologically active substances can also be present.

The compounds of the present invention may be administered by any suitable route, preferably in the form of a pharmaceutical composition adapted to such a route, and in a dose effective for the treatment intended. The active compounds and compositions, for example, may be administered orally, rectally, parenterally, or topically.

Oral administration of a solid dose form may be, for example, presented in discrete units, such as hard or soft capsules, pills, cachets, lozenges, or tablets, each containing a predetermined amount of at least one compound of the present invention. In another embodiment, the oral administration may be in a powder or granule form. In another embodiment, the oral dose form is sub-lingual, such as, for example, a lozenge. In such solid dosage forms, the compounds of Formula I are ordinarily combined with one or more adjuvants. Such capsules or tablets may contain a controlled-release formulation. In the case of capsules, tablets, and pills, the dosage forms also may comprise buffering agents or may be prepared with enteric coatings.

In another embodiment, oral administration may be in a liquid dose form. Liquid dosage forms for oral administration include, for example, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs containing inert diluents commonly used in the art (e.g., water). Such compositions also may comprise adjuvants, such as wetting, emulsifying, suspending, flavoring (e.g., sweetening), and/or perfuming agents.

In another embodiment, the present invention comprises a parenteral dose form. “Parenteral administration” includes, for example, subcutaneous injections, intravenous injections, intraperitoneal injections, intramuscular injections, intrasternal injections, and infusion. Injectable preparations (e.g., sterile injectable aqueous or oleaginous suspensions) may be formulated according to the known art using suitable dispersing, wetting agents, and/or suspending agents.

In another embodiment, the present invention comprises a topical dose form. “Topical administration” includes, for example, transdermal administration, such as via transdermal patches or iontophoresis devices, intraocular administration, or intranasal or inhalation administration. Compositions for topical administration also include, for example, topical gels, sprays, ointments, and creams. A topical formulation may include a compound which enhances absorption or penetration of the active ingredient through the skin or other affected areas. When the compounds of this invention are administered by a transdermal device, administration will be accomplished using a patch either of the reservoir and porous membrane type or of a solid matrix variety. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, dusting powders, dressings, foams, films, skin patches, wafers, implants, sponges, fibres, bandages and microemulsions. Liposomes may also be used. Typical carriers include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol and propylene glycol. Penetration enhancers may be incorporated; see, for example, J. Pharm. Sci., 88 (10), 955-958, by Finnin and Morgan (October 1999).

Formulations suitable for topical administration to the eye include, for example, eye drops wherein the compound of this invention is dissolved or suspended in a suitable carrier. A typical formulation suitable for ocular or aural administration may be in the form of drops of a micronized suspension or solution in isotonic, pH-adjusted, sterile saline. Other formulations suitable for ocular and aural administration include ointments, biodegradable (e.g., absorbable gel sponges, collagen) and non-biodegradable (e.g., silicone) implants, wafers, lenses and particulate or vesicular systems, such as niosomes or liposomes. A polymer such as cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, a cellulosic polymer, for example, hydroxypropylmethylcellulose, hydroxyethylcellulose, or methyl cellulose, or a heteropolysaccharide polymer, for example, gelan gum, may be incorporated together with a preservative, such as benzalkonium chloride. Such formulations may also be delivered by iontophoresis.

For intranasal administration or administration by inhalation, the active compounds of the invention are conveniently delivered in the form of a solution or suspension from a pump spray container that is squeezed or pumped by the patient or as an aerosol spray presentation from a pressurized container or a nebulizer, with the use of a suitable propellant. Formulations suitable for intranasal administration are typically administered in the form of a dry powder (either alone, as a mixture, for example, in a dry blend with lactose, or as a mixed component particle, for example, mixed with phospholipids, such as phosphatidylcholine) from a dry powder inhaler or as an aerosol spray from a pressurized container, pump, spray, atomizer (preferably an atomizer using electrohydrodynamics to produce a fine mist), or nebulizer, with or without the use of a suitable propellant, such as 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. For intranasal use, the powder may comprise a bioadhesive agent, for example, chitosan or cyclodextrin.

In another embodiment, the present invention comprises a rectal dose form. Such rectal dose form may be in the form of, for example, a suppository. Cocoa butter is a traditional suppository base, but various alternatives may be used as appropriate.

Other carrier materials and modes of administration known in the pharmaceutical art may also be used. Pharmaceutical compositions of the invention may be prepared by any of the well-known techniques of pharmacy, such as effective formulation and administration procedures. The above considerations in regard to effective formulations and administration procedures are well known in the art and are described in standard textbooks. Formulation of drugs is discussed in, for example, Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1975; Liberman et al., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Kibbe et al., Eds., Handbook of Pharmaceutical Excipients (3^(rd) Ed.), American Pharmaceutical Association, Washington, 1999.

The compounds of the present invention can be used, alone or in combination with other therapeutic agents, in the treatment of various conditions or disease states. The compound(s) of the present invention and other therapeutic agent(s) may be may be administered simultaneously (either in the same dosage form or in separate dosage forms) or sequentially.

Two or more compounds may be administered simultaneously, concurrently or sequentially. Additionally, simultaneous administration may be carried out by mixing the compounds prior to administration or by administering the compounds at the same point in time but at different anatomic sites or using different routes of administration.

The phrases “concurrent administration,” “co-administration,” “simultaneous administration,” and “administered simultaneously” mean that the compounds are administered in combination.

The present invention includes the use of a combination of a BACE inhibitor compound as provided in Formula I and one or more additional pharmaceutically active agent(s). If a combination of active agents is administered, then they may be administered sequentially or simultaneously, in separate dosage forms or combined in a single dosage form. Accordingly, the present invention also includes pharmaceutical compositions comprising an amount of: (a) a first agent comprising a compound of Formula I or a pharmaceutically acceptable salt of the compound; (b) a second pharmaceutically active agent; and (c) a pharmaceutically acceptable carrier, vehicle or diluent.

The compounds of this invention may also be used in conjunction with other pharmaceutical agents for the treatment of the diseases, conditions and/or disorders described herein. Therefore, methods of treatment that include administering compounds of the present invention in combination with other pharmaceutical agents are also provided. Suitable pharmaceutical agents that may be used in combination with the compounds of the present invention include, without limitation:

-   (i) anti-obesity agents (including appetite suppressants), include     gut-selective MTP inhibitors (e.g., dirlotapide, mitratapide and     implitapide, R56918 (CAS No. 403987) and CAS No. 913541-47-6), CCKa     agonists (e.g.,     N-benzyl-2-[4-(1H-indol-3-ylmethyl)-5-oxo-1-phenyl-4,5-dihydro-2,3,6,10b-tetraaza-benzo[e]azulen-6-yl]-N-isopropyl-acetamide     described in PCT Publication No. WO 2005/116034 or US Publication     No. 2005-0267100 A1), 5HT2c agonists (e.g., lorcaserin), MCR4     agonists (e.g., compounds described in U.S. Pat. No. 6,818,658),     lipase inhibitors (e.g., Cetilistat), PYY₃₋₃₆ (as used herein     “PYY₃₋₃₆” includes analogs, such as peglated PYY₃₋₃₆, e.g., those     described in US Publication 2006/0178501), opioid antagonists (e.g.,     naltrexone), oleoyl-estrone (CAS No. 180003-17-2), obinepitide     (TM30338), pramlintide (Symlin®), tesofensine (NS2330), leptin,     bromocriptine, orlistat, AOD-9604 (CAS No. 221231-10-3) and     sibutramine. -   (ii) anti-diabetic agents, such as an acetyl-CoA carboxylase (ACC)     inhibitor as described in WO2009144554, WO2003072197, WO2009144555     and WO2008065508, a diacylglycerol O-acyltransferase 1 (DGAT-1)     inhibitor, such as those described in WO09016462 or WO2010086820,     AZD7687 or LCQ908, a diacylglycerol O-acyltransferase 2 (DGAT-2)     inhibitor, a monoacylglycerol O-acyltransferase inhibitor, a     phosphodiesterase (PDE)-10 inhibitor, an AMPK activator, a     sulfonylurea (e.g., acetohexamide, chlorpropamide, diabinese,     glibenclamide, glipizide, glyburide, glimepiride, gliclazide,     glipentide, gliquidone, glisolamide, tolazamide, and tolbutamide), a     meglitinide, an α-amylase inhibitor (e.g., tendamistat, trestatin     and AL-3688), an α-glucoside hydrolase inhibitor (e.g., acarbose),     an α-glucosidase inhibitor (e.g., adiposine, camiglibose,     emiglitate, miglitol, voglibose, pradimicin-Q, and salbostatin), a     PPAR γ agonist (e.g., balaglitazone, ciglitazone, darglitazone,     englitazone, isaglitazone, pioglitazone and rosiglitazone), a PPAR     α/γ agonist (e.g., CLX-0940, GW-1536, GW-1929, GW-2433, KRP-297,     L-796449, LR-90, MK-0767 and SB-219994), a biguanide (e.g.,     metformin), a glucagon-like peptide 1 (GLP-1) modulator such as an     agonist (e.g., exendin-3 and exendin-4), liraglutide, albiglutide,     exenatide (Byetta®), albiglutide, taspoglutide, lixisenatide,     dulaglutide, semaglutide, NN-9924, TTP-054, a protein tyrosine     phosphatase-1B (PTP-1B) inhibitor (e.g., trodusquemine, hyrtiosal     extract, and compounds disclosed by Zhang, S. et al., Drug Discovery     Today, 12(9/10), 373-381 (2007)), a SIRT-1 inhibitor (e.g.,     resveratrol, GSK2245840 or GSK184072), a dipeptidyl peptidase IV     (DPP-IV) inhibitor (e.g., those in WO2005116014, sitagliptin,     vildagliptin, alogliptin, dutogliptin, linagliptin and saxagliptin),     an insulin secretagogue, a fatty acid oxidation inhibitor, an A2     antagonist, a c-jun amino-terminal kinase (JNK) inhibitor, a     glucokinase activator (GKa) such as those described in WO2010103437,     WO2010103438, WO2010013161, WO2007122482, TTP-399, TTP-355, TTP-547,     AZD1656, ARRY403, MK-0599, TAK-329, AZD5658 or GKM-001, insulin, an     insulin mimetic, a glycogen phosphorylase inhibitor (e.g.,     GSK1362885), a VPAC2 receptor agonist, an SGLT2 inhibitor, such as     those described in E. C. Chao et al., Nature Reviews Drug Discovery     9, 551-559 (July 2010) including dapagliflozin, canagliflozin,     BI-10733, tofogliflozin (CSG452), ASP-1941, THR1474, TS-071,     ISIS388626 and LX4211 as well as those in WO2010023594, a glucagon     receptor modulator such as those described in Demong, D. E. et al.,     Annual Reports in Medicinal Chemistry 2008, 43, 119-137, a GPR119     modulator, particularly an agonist, such as those described in     WO2010140092, WO2010128425, WO2010128414, WO2010106457, Jones, R. M.     et al., in Medicinal Chemistry 2009, 44, 149-170 (e.g. MBX-2982,     GSK1292263, APD597 and PSN821), an FGF21 derivative or an analog     such as those described in Kharitonenkov, A. et al., Current Opinion     in Investigational Drugs 2009, 10(4), 359-364, TGR5 (also termed     GPBAR1) receptor modulators, particularly agonists, such as those     described in Zhong, M., Current Topics in Medicinal Chemistry, 2010,     10(4), 386-396 and INT777, a GPR40 agonist, such as those described     in Medina, J.C., Annual Reports in Medicinal Chemistry, 2008, 43,     75-85, including but not limited to TAK-875, a GPR120 modulator,     particularly an agonist, a high affinity nicotinic acid receptor     (HM74A) activator, and an SGLT1 inhibitor, such as GSK1614235. A     further representative listing of anti-diabetic agents that can be     combined with the compounds of the present invention can be found,     for example, at page 28, line 35 through page 30, line 19 of     WO2011005611. Preferred anti-diabetic agents are metformin and     DPP-IV inhibitors (e.g., sitagliptin, vildagliptin, alogliptin,     dutogliptin, linagliptin and saxagliptin). Other antidiabetic agents     could include inhibitors or modulators of carnitine palmitoyl     transferase enzymes, inhibitors of fructose 1,6-diphosphatase,     inhibitors of aldose reductase, mineralocorticoid receptor     inhibitors, inhibitors of TORC2, inhibitors of CCR2 and/or CCR5,     inhibitors of PKC isoforms (e.g., PKCa, PKCb, PKCg), inhibitors of     fatty acid synthetase, inhibitors of serine palmitoyl transferase,     modulators of GPR81, GPR39, GPR43, GPR41, GPR105, Kv1.3, retinol     binding protein 4, glucocorticoid receptor, somatostain receptors     (e.g. SSTR1, SSTR2, SSTR3 and SSTR5), inhibitors or modulators of     PDHK2 or PDHK4, inhibitors of MAP4K4, modulators of IL1 family     including IL1beta, and modulators of RXRalpha. In addition, suitable     anti-diabetic agents include mechanisms listed by Carpino, P. A.,     Goodwin, B. Expert Opin. Ther. Pat, 2010, 20(12), 1627-51; -   (iii) anti-hyperglycemic agents, for example, those described at     page 31, line 31 through page 32, line 18 of WO 2011005611; -   (iv) lipid lowering agents (for example, those described at page 30,     line 20 through page 31, line 30 of WO 2011005611), and     anti-hypertensive agents (for example, those described at page 31,     line 31 through page 32, line 18 of WO 2011005611); -   (v) acetylcholinesterase inhibitors, such as donepezil hydrochloride     (ARICEPT®, MEMAC), physostigmine salicylate (ANTILIRIUM®),     physostigmine sulfate (ESERINE), ganstigmine, rivastigmine     (EXELON®), ladostigil, NP-0361, galantamine hydrobromide (RAZADYNE®,     REMINYL®, NIVALIN®), tacrine (COGNEX®), tolserine, memoquin,     huperzine A (HUP-A; Neuro-Hitech), phenserine, bisnorcymserine (also     known as BNC), and INM-176; -   (vi) amyloid-β (or fragments thereof), such as Aβ₁₋₁₅ conjugated to     pan HLA DR-binding epitope (PADRE®), ACC-001 (Elan/Wyeth), and     Affitope; -   (vii) antibodies to amyloid-β (or fragments thereof), such as     ponezumab, solanezumab, bapineuzumab (also known as AAB-001),     AAB-002 (Wyeth/Elan), Gantenerumab, intravenous Ig (GAMMAGARD®),     LY2062430 (humanized m266; Lilly), and those disclosed in     International Patent Publication Nos WO04/032868, WO05/025616,     WO06/036291, WO06/069081, WO06/118959, in US Patent Publication Nos     US2003/0073655, US2004/0192898, US2005/0048049, US2005/0019328, in     European Patent Publication Nos EP0994728 and 1257584, and in U.S.     Pat. No. 5,750,349; -   (viii) amyloid-lowering or -inhibiting agents (including those that     reduce amyloid production, accumulation and fibrillization) such as     eprodisate, celecoxib, lovastatin, anapsos, colostrinin,     pioglitazone, clioquinol (also known as PBT1), PBT2 (Prana     Biotechnology), flurbiprofen (ANSAID®, FROBEN®) and its R-enantiomer     tarenflurbil (FLURIZAN®), nitroflurbiprofen, fenoprofen (FENOPRON,     NALFON®), ibuprofen (ADVIL®, MOTRIN®, NUROFEN®), ibuprofen lysinate,     meclofenamic acid, meclofenamate sodium (MECLOMEN®), indomethacin     (INDOCIN®), diclofenac sodium (VOLTAREN®), diclofenac potassium,     sulindac (CLINORIL®), sulindac sulfide, diflunisal (DOLOBID®),     naproxen (NAPROSYN®), naproxen sodium (ANAPROX®, ALEVE®),     insulin-degrading enzyme (also known as insulysin), the gingko     biloba extract EGb-761 (ROKAN®, TEBONIN®), tramiprosate (CEREBRIL®,     ALZHEMED®), KIACTA®), neprilysin (also known as neutral     endopeptidase (NEP)), scyllo-inositol (also known as scyllitol),     atorvastatin (LIPITOR®), simvastatin (ZOCOR®), ibutamoren mesylate,     BACE inhibitors such as LY450139 (Lilly), BMS-782450, GSK-188909;     Gamma Secretase Modulators and Inhibitors such as ELND-007,     BMS-708163 (Avagacestat), and DSP8658 (Dainippon); and RAGE     (receptor for advanced glycation end-products) inhibitors, such as     TTP488 (Transtech) and TTP4000 (Transtech), and those disclosed in     U.S. Pat. No. 7,285,293, including PTI-777; -   (ix) alpha-adrenergic receptor agonists, and beta-adrenergic     receptor blocking agents (beta blockers); anticholinergics;     anticonvulsants; antipsychotics; calcium channel blockers; catechol     O-methyltransferase (COMT) inhibitors; central nervous system     stimulants; corticosteroids; dopamine receptor agonists and     antagonists; dopamine reuptake inhibitors; gamma-aminobutyric acid     (GABA) receptor agonists; immunosuppressants; interferons;     muscarinic receptor agonists; neuroprotective drugs; nicotinic     receptor agonists; norepinephrine (noradrenaline) reuptake     inhibitors; quinolines; and trophic factors; -   (x) histamine 3 (H3) antagonists, such as PF-3654746 and those     disclosed in US Patent Publication Nos US2005-0043354,     US2005-0267095, US2005-0256135, US2008-0096955, US2007-1079175, and     US2008-0176925; International Patent Publication Nos WO2006/136924,     WO2007/063385, WO2007/069053, WO2007/088450, WO2007/099423,     WO2007/105053, WO2007/138431, and WO2007/088462; and U.S. Pat. No.     7,115,600); -   (xi) N-methyl-D-aspartate (NMDA) receptor antagonists, such as     memantine (NAMENDA, AXURA, EBIXA), amantadine (SYMMETREL),     acamprosate (CAMPRAL), besonprodil, ketamine (KETALAR), delucemine,     dexanabinol, dexefaroxan, dextromethorphan, dextrorphan,     traxoprodil, CP-283097, himantane, idantadol, ipenoxazone, L-701252     (Merck), lancicemine, levorphanol (DROMORAN), methadone,     (DOLOPHINE), neramexane, perzinfotel, phencyclidine, tianeptine     (STABLON), dizocilpine (also known as MK-801), ibogaine, voacangine,     tiletamine, riluzole (RILUTEK), aptiganel (CERESTAT), gavestinel,     and remacimide; -   (xii) monoamine oxidase (MAO) inhibitors, such as selegiline     (EMSAM), selegiline hydrochloride (1-deprenyl, ELDEPRYL, ZELAPAR),     dimethylselegiline, brofaromine, phenelzine (NARDIL),     tranylcypromine (PARNATE), moclobemide (AURORIX, MANERIX),     befloxatone, safinamide, isocarboxazid (MARPLAN), nialamide     (NIAMID), rasagiline (AZILECT), iproniazide (MARSILID, IPROZID,     IPRONID), iproclozide, toloxatone (HUMORYL, PERENUM), bifemelane,     desoxypeganine, harmine (also known as telepathine or banasterine),     harmaline, linezolid (ZYVOX, ZYVOXID), and pargyline (EUDATIN,     SUPIRDYL); -   (xiii) phosphodiesterase (PDE) inhibitors, including (a) PDE1     inhibitors (b) PDE2 inhibitors (c) PDE3 inhibitors (d) PDE4     inhibitors (e) PDE5 inhibitors (f) PDE9 inhibitors (e.g.,     PF-04447943, BAY 73-6691 (Bayer AG) and those disclosed in US Patent     Publication Nos US2003/0195205, US2004/0220186, US2006/0111372,     US2006/0106035, and U.S. Ser. No. 12/118,062 (filed May 9, 2008)),     and (g) PDE10 inhibitors such as     2-({4-[1-methyl-4-(pyridin-4-yl)-1H-pyrazol-3-yl]phenoxy}methyl)quinoline     (PF-2545920); -   (xiv) serotonin (5-hydroxytryptamine) 1A (5-HT_(1A)) receptor     antagonists, such as spiperone, levo-pindolol, lecozotan; -   (xv) serotonin (5-hydroxytryptamine) 2C (5-HT_(2c)) receptor     agonists, such as vabicaserin, and zicronapine; serotonin     (5-hydroxytryptamine) 4 (5-HT₄) receptor agonists/antagonists, such     as PRX-03140 (Epix) and PF-04995274; -   (xvi) serotonin (5-hydroxytryptamine) 3C (5-HT_(3c)) receptor     antagonists, such as Ondansetron (Zofran); -   (xvii) serotonin (5-hydroxytryptamine) 6 (5-HT₆) receptor     antagonists, such as mianserin (TOLVON, BOLVIDON, NORVAL),     methiothepin (also known as metitepine), ritanserin, SB-271046,     SB-742457 (GlaxoSmithKline), Lu AE58054 (Lundbeck A/S), SAM-760, and     PRX-07034 (Epix); -   (xviii) serotonin (5-HT) reuptake inhibitors such as alaproclate,     citalopram (CELEXA, CIPRAMIL), escitalopram (LEXAPRO, CIPRALEX),     clomipramine (ANAFRANIL), duloxetine (CYMBALTA), femoxetine     (MALEXIL), fenfluramine (PONDIMIN), norfenfluramine, fluoxetine     (PROZAC), fluvoxamine (LUVOX), indalpine, milnacipran (IXEL),     paroxetine (PAXIL, SEROXAT), sertraline (ZOLOFT, LUSTRAL), trazodone     (DESYREL, MOLIPAXIN), venlafaxine (EFFEXOR), zimelidine (NORMUD,     ZELMID), bicifadine, desvenlafaxine (PRISTIQ), brasofensine,     vilazodone, cariprazine and tesofensine; -   (xix) Glycine transporter-1 inhibitors such as paliflutine,     ORG-25935, and ORG-26041; and nnGluR modulators such as AFQ-059 and     amantidine; -   (xx) AMPA-type glutamate receptor modulators such as perampanel,     mibampator, selurannpanel, GSK-729327, and     N-{(3S,4S)-4-[4-(5-cyanothiophen-2-yl)phenoxy]tetrahydrofuran-3-yl}propane-2-sulfonamide; -   (xxi) P450 inhibitors, such as ritonavir; -   (xxii) tau therapy targets, such as davunetide;

and the like.

The present invention further comprises kits that are suitable for use in performing the methods of treatment described above. In one embodiment, the kit contains a first dosage form comprising one or more of the compounds of the present invention and a container for the dosage, in quantities sufficient to carry out the methods of the present invention.

In another embodiment, the kit of the present invention comprises one or more compounds of the invention.

General Synthetic Schemes

The compounds of Formula I may be prepared by the methods described below, together with synthetic methods known in the art of organic chemistry, or modifications and transformations that are familiar to those of ordinary skill in the art. The starting materials used herein are commercially available or may be prepared by routine methods known in the art [such as those methods disclosed in standard reference books such as the Compendium of Organic Synthetic Methods, Vol. I-XII (published by Wiley-Interscience)]. Preferred methods include, but are not limited to, those described below.

During any of the following synthetic sequences it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This can be achieved by means of conventional protecting groups, such as those described in T. W. Greene, Protective Groups in Organic Chemistry, John Wiley & Sons, 1981; T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Chemistry, John Wiley & Sons, 1991; and T. W. Greene and P. G. M. Wuts, Protective Groups in Organic Chemistry, John Wiley & Sons, 1999, which are hereby incorporated by reference.

Compounds of Formula I, or their pharmaceutically acceptable salts, can be prepared according to the reaction Schemes discussed herein below. Unless otherwise indicated, the substituents in the Schemes are defined as above. Isolation and purification of the products is accomplished by standard procedures, which are known to a chemist of ordinary skill.

One skilled in the art will recognize that in many cases, the compounds in Schemes 1 through 11 will be generated as a mixture of diastereomers and/or enantiomers; these may be separated at various stages of the synthetic schemes using conventional techniques or a combination of such techniques, such as, but not limited to, crystallization, normal-phase chromatography, reversed phase chromatography and chiral chromatography, to afford the single enantiomers of the invention.

It will be understood by one skilled in the art that the various symbols, superscripts and subscripts used in the schemes, methods and examples are used for convenience of representation and/or to reflect the order in which they are introduced in the schemes, and are not intended to necessarily correspond to the symbols, superscripts or subscripts in the appended claims. The schemes are representative of methods useful in synthesizing the compounds of the present invention. They are not to constrain the scope of the invention in any way.

Scheme 1 refers to the preparation of compounds of Formula I. Referring to Scheme 1, the compound of Formula I can be prepared from the compound of Formula II through a removal of protecting group P¹. P¹ in this case refers to groups well known to those skilled in the art for amine protection. For example, P¹ may be a benzoyl group (Bz), which can be cleaved via acidic conditions, or through treatment with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) in methanol. Alternatively P¹ may be one of many protecting group suitable for amines, including 9-fluorenylnnethoxycarbonyl (Fmoc) or tert-butoxycarbonyl (BOC) and can be cleaved under standard conditions known to one skilled in the art.

Scheme 2 refers to the preparation of compounds II wherein P¹ is Bz or Fmoc. The oxidation of compounds of Formula III can be accomplished by a number of standard oxidation protocols, for instance using tetrapropylammonium perruthenate (TPAP) and N-methylmorpholine-N-oxide (NMO) in acetonitrile. Carboxylic acid IV can be converted to compounds of Formula II via a number of methods outlined in the following reference: Practical Synthetic Organic Chemistry: Reactions, Principles, and Techniques. 2011, Chapter 13, Wiley & Sons, Inc., Caron, S., ed., as well as additional methods known to those skilled in the art. Compound II can be converted into a compound of Formula I according to the methods of Scheme 1.

Scheme 3 refers to the preparation of compounds II wherein P¹ is Bz or Fmoc. The preparation of compounds of Formula V can be effected by activation of the acid IV using a standard peptide coupling reagent, for instance using 2-[2-oxo-1(2H)-pyridyl]-1,1,3,3-tetramethyluronium tetrafluoroborate (TPTU), followed by treatment with an appropriate ammonia source, for instance, a solution of ammonia in dioxane. Amides of Formula V can be converted to compounds of Formula II via a number of methods outlined in the following reference: Practical Synthetic Organic Chemistry: Reactions, Principles, and Techniques. 2011, Chapter 13, Wiley & Sons, Inc., Caron, S., ed. Compound II can be converted into a compound of Formula I according to the methods of Scheme 1.

Scheme 4 refers to the preparation of compounds II wherein P¹ is Bz or Fmoc. The oxidation of compounds of Formula III can be effected by a number of standard oxidation protocols, for instance using Dess-Martin periodinane or sulfur trioxide-pyridine with DMSO (Parikh-Doering conditions). Aldehyde VI can be converted to compounds of Formula II via a number of methods outlined in the following reference: Practical Synthetic Organic Chemistry: Reactions, Principles, and Techniques. 2011, Chapter 13, Wiley & Sons, Inc., Caron, S., ed. Compound II can be converted into a compound of Formula I according to the methods of Scheme 1.

Scheme 5 refers to the preparation of compounds II wherein P¹ is Bz or Fmoc. Compounds of Formula VII can be prepared by treatment of amide V with a suitable methylating agent, for instance trimethyloxonium tetrafluoroborate, followed by treatment with an ammonia source, for instance a solution of ammonia in methanol. Amidine VII can be converted to compounds of Formula II via a number of methods outlined in the following reference: Practical Synthetic Organic Chemistry: Reactions, Principles, and Techniques. 2011, Chapter 13, Wiley & Sons, Inc., Caron, S., ed. Compound II can be converted into a compound of Formula I according to the methods of Scheme 1.

Scheme 6 refers to the preparation of compounds II wherein P¹ is Bz or Boc. The addition of an organometallic derivative (magnesiate or lithiate) of R² to compounds of Formula VIII under standard anionic conditions, for instance in tetrahydrofuran (THF) at −78° C., provides compounds of Formula IX. Subsequent reduction of the resultant lactol using standard reduction conditions, for instance trimethylsilyl trifluoromethanesulfonate (TMSOTf) and triethylsilane, provides compounds of Formula II. Compound II can be converted into a compound of Formula I according to the methods of Scheme 1.

Scheme 7 refers to the preparation of compounds II wherein P¹ is Bz or Fmoc. The treatment of lactones of Formula VIII with base, for instance potassium hexamethyldisilazide (KHMDS), and N-(5-chloro-2-pyridyl)bis(trifluoronnethanesulfoninnide (Comins' Reagent) provides compounds of Formula X. The reaction of enol triflate X with the corresponding R²-containing boronic acid using standard Suzuki reaction conditions replaces the triflate with R²; subsequent reduction of the resultant enol using standard reduction conditions, for instance trimethylsilyl trifluoromethanesulfonate (TMSOTf) and triethylsilane provides compounds of Formula II. Compound II can be converted into a compound of Formula I according to the methods of Scheme 1.

Scheme 8 refers to the preparation of compounds III wherein P¹ is Bz or Fmoc. Isoxazolidines of Formula XI (which may be obtained via the chemistry depicted in Scheme 11, utilizing a benzyloxymethyl group in place of R²) are subjected to reducing conditions, for instance zinc in acetic acid, affording compounds of Formula XII. The amino alcohols XII are treated with an isothiocyanate, for instance benzoyl isothiocyanate, to provide thioureas of Formula XIII. Cyclization is induced using strong acid, including for instance sulfuric acid, or alternatively, standard Mitsunobu conditions, to give compounds of Formula XIV. Cleavage of the benzyl ether under standard conditions, for instance using boron trichloride, provides alcohols of Formula III. Compound III can be converted into a compound of Formula II according to the methods of Schemes 2 or 4.

Scheme 9 refers to the preparation of compounds of Formula VIII wherein P¹ is Bz or Fmoc. Aldehydes of Formula VI are subjected to basic conditions, for instance potassium carbonate in acetonitrile, and trapped using an appropriate anhydride, for instance acetic anhydride, to afford protected enols of Formula XV, wherein P² is an acyl group. Oxidative cleavage of the resulting enol moiety using standard conditions, including for instance, ruthenium chloride and sodium periodate, affords lactones of Formula VIII. Compound VIII can be converted into a compound of Formula I according to the methods of Scheme 6 and 7.

Scheme 10 refers to the preparation of compounds II wherein P¹ is Bz or Fmoc. Isoxazolidines of Formula XVI are subjected to reducing conditions, for instance zinc in acetic acid, affording compounds of Formula XVII. The resulting amino alcohols are treated with an isothiocyanate, for instance benzoyl isothiocyanate, to provide thioureas of Formula XVIII. Cyclization is induced using strong acid, including for instance, sulfuric acid, or alternatively, standard Mitsunobu conditions, to give compounds of Formula II. Compound II can be directly converted into a compound of Formula I according to the methods of Scheme 1.

Scheme 11 refers to the preparation of compound XVI. Homoallylic alcohol XIX is alkylated with 2-bromo-1,1-dimethoxyethane under basic conditions, such as treatment with potassium hydride, to provide the corresponding ether XX. The acetal is cleaved under acidic conditions, aqueous HCl as an example, to give aldehyde XXI. Condensation with a hydroxylamine salt, such as hydroxylamine sulfate, provides a geometric mixture of the corresponding oxime XXII. Cycloaddition to form isoxazoline XXIII may be carried out by treatment of oxime XXII with an oxidizing agent, such as sodium hypochlorite or N-chlorosuccinimide. Reaction of isoxazoline) XXIII with an appropriate arylmetallic reagent (for instance, an aryllithium such as 2,4-difluorophenyllithium, or the corresponding aryl Grignard reagent) at low temperature, e.g., −78° C., yields compounds of Formula XVI. One of ordinary skill in the art will recognize that the stereochemistry of addition of the arylmetallic reagent is determined by the stereochemistry of the adjacent methine center, yielding a racemic mixture of cis-fused diastereomers, which can be converted into compounds of Formula I according to the methods of Schemes 10 and 1.

Experimental Procedures and Working Examples

The following illustrate the synthesis of various compounds of the present invention. Additional compounds within the scope of this invention may be prepared using the methods illustrated in these Examples, either alone or in combination with techniques generally known in the art.

Experiments were generally carried out under inert atmosphere (nitrogen or argon), particularly in cases where oxygen- or moisture-sensitive reagents or intermediates were employed. Commercial solvents and reagents were generally used without further purification, including anhydrous solvents where appropriate (generally Sure-Seal™ products from the Aldrich Chemical Company, Milwaukee, Wis.). Products were generally dried under vacuum before being carried on to further reactions or submitted for biological testing. Mass spectrometry data is reported from either liquid chromatography-mass spectrometry (LCMS), atmospheric pressure chemical ionization (APCI) or gas chromatography-mass spectrometry (GCMS) instrumentation. Chemical shifts for nuclear magnetic resonance (NMR) data are expressed in parts per million (ppm, δ) referenced to residual peaks from the deuterated solvents employed.

For syntheses referencing procedures in other Examples or Methods, reaction conditions (length of reaction and temperature) may vary. In general, reactions were followed by thin layer chromatography or mass spectrometry, and subjected to work-up when appropriate. Purifications may vary between experiments: in general, solvents and the solvent ratios used for eluents/gradients were chosen to provide appropriate R_(f)s or retention times.

Preparation P1 N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(hydroxymethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (P1)

Step 1. Synthesis of (2R)-1-(benzyloxy)pent-4-en-2-ol (C1)

To a solution of (2R)-2-[(benzyloxy)methyl]oxirane (167 g, 1.02 mol) in tetrahydrofuran (2 L) was added copper(I) iodide (11.62 g, 61.02 mmol) at room temperature. The mixture was stirred for 5 minutes, then cooled to 78° C. A solution of vinylmagnesium bromide (1 M in tetrahydrofuran, 1.12 L, 1.12 mol) was added drop-wise over 1 hour while the reaction temperature was maintained below 70° C. Upon completion of the addition, the cooling bath was removed and the reaction mixture was left to stir at room temperature for 1 hour, then quenched by slow addition of aqueous ammonium chloride solution (200 mL). After dilution with aqueous ammonium chloride solution (1.5 L) and ethyl acetate (1.5 L), the aqueous layer was extracted with ethyl acetate (1 L) and the combined organic layers were washed with aqueous ammonium chloride solution (1.5 L), dried over magnesium sulfate, filtered, and concentrated in vacuo. Three batches of this reaction were carried out and combined to give the product as an orange oil. Yield: 600 g, 3.1 mol, quantitative. ¹H NMR (400 MHz, CDCl₃) δ 7.28-7.40 (m, 5H), 5.78-5.90 (m, 1H), 5.08-5.17 (m, 2H), 4.57 (s, 2H), 3.86-3.94 (m, 1H), 3.53 (dd, J=9.6, 3.3 Hz, 1H), 3.39 (dd, J=9.6, 7.4 Hz, 1H), 2.26-2.34 (m, 3H).

Step 2. Synthesis of ({[(2R)-2-(2,2-diethoxyethoxy)pent-4-en-1-yl]oxy}methyl)benzene (C2)

To a suspension of sodium hydride (60% in mineral oil, 98.8 g, 2.47 mol) in tetrahydrofuran (1 L) at room temperature was added drop-wise over 30 minutes a solution of (2R)-1-(benzyloxy)pent-4-en-2-ol (C1) (190 g, 0.988 mol) in tetrahydrofuran (500 mL), while the reaction temperature was maintained below 30° C. After 30 minutes, a solution of 2-bromo-1,1-diethoxyethane (390 g, 1.98 mol) in tetrahydrofuran (500 mL) was added drop-wise. The reaction mixture was stirred at room temperature for 1 hour, then the temperature was gradually increased to 70° C. and the reaction mixture was left to stir at 70° C. for 18 hours. It was then cooled to room temperature, subsequently cooled in an ice bath, and quenched by slow addition of ice/water (200 mL), while keeping the internal reaction temperature at approximately 18° C. The mixture was diluted with saturated aqueous sodium chloride solution (1 L) and ethyl acetate (1 L), and the organic layer was washed with saturated aqueous sodium chloride solution (1 L), dried over magnesium sulfate, filtered, and concentrated under reduced pressure. Purification was effected by filtration through a pad of silica (Gradient: 0% to 20% ethyl acetate in heptane) to afford the product as an orange oil. Yield: 257 g of 60% purity, approximately 500 mmol, 51% yield and 57.76 g of 90% purity, approximately 170 mmol, 17% yield. ¹H NMR (400 MHz, CDCl₃), product peaks only: δ 7.26-7.38 (m, 5H), 5.78-5.90 (m, 1H), 5.02-5.13 (m, 2H), 4.61 (t, J=5.3 Hz, 1H), 4.55 (s, 2H), 3.48-3.74 (m, 9H), 2.31-2.37 (m, 2H), 1.22 (t, J=7.1 Hz, 3H), 1.21 (t, J=7.1 Hz, 3H).

Step 3. Synthesis of 2-{[(2R)-1-(benzyloxy)pent-4-en-2-yl]oxy}-N-hydroxyethanimine (C3)

A solution of ({[(2R)-2-(2,2-diethoxyethoxy)pent-4-en-1-yl]oxy}methyl)benzene (C2) (234 g, 0.759 mol) in formic acid (400 mL) and water (100 mL) was stirred at room temperature for 2 hours. As LCMS analysis revealed a small amount of remaining starting material, formic acid (50 mL) was added and the reaction mixture was stirred for a further 30 minutes. The reaction mixture was diluted with ethanol (1 L) and water (400 mL). Hydroxylamine sulfate (435 g, 2.65 mol) and sodium acetate (217 g, 2.64 mol) were added and the reaction was stirred at room temperature for 18 hours. The reaction mixture was then filtered and concentrated in vacuo. The residue was partitioned between ethyl acetate (500 mL) and water (1 L), and the aqueous layer was extracted with ethyl acetate (3×500 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (2×500 mL), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to provide the product as an orange oil. By ¹H NMR, this material consisted of a roughly 1:1 mixture of oxime isomers. Yield: 234 g, which was taken directly to the following step. LCMS m/z 250.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ [7.52 (t, J=5.5 Hz) and 6.96 (t, J=3.6 Hz), total 1H], 7.28-7.39 (m, 5H), 5.74-5.87 (m, 1H), 5.04-5.14 (m, 2H), 4.55 and 4.56 (2 s, total 2H), {4.45-4.55 (m) and [4.27 (dd, half of ABX pattern, J=13.2, 5.4 Hz) and 4.21 (dd, half of ABX pattern, J=13.2, 5.6 Hz)], total 2H}, 2.30-2.37 (m, 2H).

Step 4. Synthesis of (3aR,5R)-5-[(benzyloxy)methyl]-3,3a,4,5-tetrahydro-7H-pyrano[3,4-c][1,2]oxazole (C4)

An aqueous solution of sodium hypochlorite (14.5% solution, 600 mL) was added drop-wise to a 0° C. solution of 2-{[(2R)-1-(benzyloxy)pent-4-en-2-yl]oxy}-N-hydroxyethanimine (C3) (224 g from the previous step, ≦0.759 mol) in dichloromethane (1 L), while the internal temperature was maintained below 15° C. After completion of the addition, the reaction mixture was left to stir at 0° C. for 1.5 hours, then diluted with water (1 L) and dichloromethane (500 mL). The aqueous layer was extracted with dichloromethane (2×500 mL), and the combined organic layers were washed with saturated aqueous sodium chloride solution (500 mL), water (500 mL) and again with saturated aqueous sodium chloride solution (500 mL). They were subsequently dried over magnesium sulfate, filtered, and concentrated in vacuo. Purification via silica gel chromatography (Gradient: 0% to 25% ethyl acetate in heptane) afforded the product as a colorless oil. The indicated relative stereochemistry of compound C4 was assigned based on nuclear Overhauser enhancement studies, which revealed an interaction between the methine protons on carbons 3a and 5. Yield: 85.3 g, 345 mmol, 45% over 2 steps. LCMS m/z 248.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.27-7.40 (m, 5H), 4.77 (d, J=13.5 Hz, 1H), 4.54-4.65 (m, 3H), 4.22 (dd, J=13.5, 1 Hz, 1H), 3.79 (dd, J=11.7, 8.0 Hz, 1H), 3.69-3.76 (m, 1H), 3.57 (dd, half of ABX pattern, J=10.1, 5.9 Hz, 1H), 3.49 (dd, half of ABX pattern, J=10.1, 4.3 Hz, 1H), 3.39-3.5 (m, 1H), 2.20 (ddd, J=12.9, 6.5, 1.6 Hz, 1H), 1.51-1.62 (m, 1H).

Step 5. Synthesis of (3aR,5R,7aS)-5-[(benzyloxy)methyl]-7a-(2,4-difluorophenyl)hexahydro-1H-pyrano[3,4-c][1,2]oxazole (C5)

Boron trifluoride diethyl etherate (60.1 mL, 474 mmol) was added to a solution of (3aR,5R)-5-[(benzyloxy)methyl]-3,3a,4,5-tetrahydro-7H-pyrano[3,4-c][1,2]oxazole (C4) (50.0 g, 202 mmol) in a 1:1 mixture of toluene and diisopropyl ether (2 L) at an internal temperature of −76° C. The reaction was stirred at this temperature for 30 minutes, then treated with 2,4-difluoro-1-iodobenzene (27.1 mL, 226 mmol). While the reaction temperature was maintained at −76 to −71° C., n-butyllithium (2.5 M in hexanes, 85.7 mL, 214 mmol) was slowly added. The reaction mixture was stirred at −76° C. for 1.5 hours, then was quenched with saturated aqueous ammonium chloride solution (1 L) and partitioned between water (1 L) and ethyl acetate (750 mL). After the heterogeneous mixture warmed to room temperature, the aqueous layer was extracted with ethyl acetate (3×250 mL), and the combined organic layers were washed with saturated aqueous sodium chloride solution (550 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Chromatography on silica gel (Gradient: 0% to 70% ethyl acetate in heptane) afforded the product as a yellow oil. Yield: 48.14 g, 133.2 mmol, 66%. ¹H NMR (400 MHz, CDCl₃) δ 7.94 (ddd, J=9, 9, 7 Hz, 1H), 7.28-7.40 (m, 5H), 6.87-6.93 (m, 1H), 6.80 (ddd, J=12.0, 8.6, 2.4 Hz, 1H), 4.60 (AB quartet, J_(AB)=12.1 Hz, Δν_(AB)=21.4 Hz, 2H), 4.14 (br dd, J=12.8, 1.3 Hz, 1H), 3.82-3.90 (m, 2H), 3.72 (d, J=7.2 Hz, 1H), 3.54-3.60 (m, 2H), 3.50 (dd, half of ABX pattern, J=10.3, 4.1 Hz, 1H), 3.04-3.13 (m, 1H), 1.86 (ddd, J=14.0, 7.0, 2.0 Hz, 1H), 1.49-1.61 (m, 1H).

Step 6. Synthesis of [(2R,4R,5S)-5-amino-2-[(benzyloxy)methyl]-5-(2,4-difluorophenyl)tetrahydro-2H-pyran-4-yl]methanol (C6)

(3aR,5R,7aS)-5-[(Benzyloxy)methyl]-7a-(2,4-difluorophenyl)hexahydro-1H-pyrano[3,4-c][1,2]oxazole (C5) (48.1 g, 133 mmol) was dissolved in acetic acid (444 mL) and treated with zinc powder (113 g, 1.73 mol). The reaction mixture, which had warmed to 40° C., was allowed to cool to room temperature and stir for 16 hours. Insoluble material was removed via filtration through a Celite pad, and the pad was washed with ethyl acetate (3×500 mL). The combined filtrates were neutralized with saturated aqueous sodium bicarbonate solution (2.5 L), and the aqueous layer was extracted with ethyl acetate (3×500 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (1 L), dried over sodium sulfate, filtered, and concentrated under reduced pressure to provide the product as a thick yellow oil, which was used in the following reaction without additional purification. Yield: 48.7 g, assumed quantitative. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ 7.62-7.80 (br m, 1H), 7.28-7.39 (m, 5H), 6.94-7.06 (m, 1H), 6.83 (ddd, J=12.7, 8.5, 2.6 Hz, 1H), 4.61 (AB quartet, upfield doublet is broadened, J_(AB)=12.2 Hz, Δν_(AB)=30.5 Hz, 2H), 4.22 (dd, J=11.6, 2.2 Hz, 1H), 3.83-3.92 (br m, 1H), 3.62-3.73 (br m, 1H), 3.56 (dd, J=10.2, 3.5 Hz, 1H), 3.34-3.41 (m, 1H), 2.26-2.43 (br m, 1H), 2.00-2.17 (br m, 1H), 1.65 (ddd, J=14.1, 4.5, 2.5 Hz, 1H).

Step 7. Synthesis of N-{[(3S,4R,6R)-6-[(benzyloxy)methyl]-3-(2,4-difluorophenyl)-4-(hydroxymethyl)tetrahydro-2H-pyran-3-yl]carbamothioyl}benzamide (C7)

Benzoyl isothiocyanate (17.8 mL, 132 mmol) was added to a solution of [(2R,4R,5S)-5-amino-2-[(benzyloxy)methyl]-5-(2,4-difluorophenyl)tetrahydro-2H-pyran-4-yl]methanol (C6) (48.7 g, 134 mmol) in dichloromethane (1.34 L), and the reaction mixture was allowed to stir at room temperature for 18 hours. Removal of solvent in vacuo afforded the product as a white solid, which was used without additional purification. Yield: 72.2 g, assumed quantitative. LCMS m/z 527.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD), characteristic peaks: δ 7.89-7.93 (m, 2H), 7.62-7.67 (m, 1H), 7.50-7.56 (m, 2H), 7.42-7.54 (br m, 1H), 7.31-7.36 (m, 2H), 7.17-7.28 (m, 3H), 6.86-6.98 (m, 2H), 4.57 (AB quartet, J_(AB)=11.9 Hz, Δν_(AB)=11.8 Hz, 2H), 3.84-3.91 (m, 1H), 3.64 (br dd, half of ABX pattern, J=10.6, 6.0 Hz, 1H), 3.58 (dd, half of ABX pattern, J=10.6, 3.8 Hz, 1H), 3.44-3.54 (br m, 1H), 2.32-2.59 (br m, 1H), 1.82-2.06 (m, 2H).

Step 8. Synthesis of N-[(4aR,6R,8aS)-6-[(benzyloxy)methyl]-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C8)

Pyridine (11.0 mL, 137 mmol) was added to a solution of N-{[(3S,4R,6R)-6-[(benzyloxy)methyl]-3-(2,4-difluorophenyl)-4-(hydroxymethyl)tetrahydro-2H-pyran-3-yl]carbamothioyl}benzamide (C7) (19.00 g, 36.08 mmol) in dichloromethane (150 mL), and the resulting solution was cooled to −50 to −60° C. Trifluoromethanesulfonic anhydride (12.1 mL, 71.9 mmol) in dichloromethane (50 mL) was added drop-wise, and the reaction mixture was gradually warmed to −5° C. over 3 hours. Water was added, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered, and concentrated in vacuo. Purification via silica gel chromatography (Gradient: 20% to 40% ethyl acetate in heptane) provided the product as a yellow foam. Yield: 15.51 g, 30.50 mmol, 85%. LCMS m/z 509.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.23 (br d, J=7 Hz, 2H), 7.37-7.57 (br m, 4H), 7.24-7.36 (m, 5H), 6.85-6.97 (m, 2H), 4.58 (AB quartet, upfield signals are slightly broadened, J_(AB)=11.9 Hz, Δν_(AB)=23.5 Hz, 2H), 4.17 (br d, J=12 Hz, 1H), 3.90-3.97 (m, 1H), 3.83 (br d, J=12 Hz, 1H), 3.64 (dd, half of ABX pattern, J=10.1, 6.4 Hz, 1H), 3.50 (dd, half of ABX pattern, J=10.2, 4.4 Hz, 1H), 3.11-3.21 (br m, 1H), 3.02 (dd, J=12.9, 4.1 Hz, 1H), 2.64 (br d, J=13 Hz, 1H), 1.92-2.05 (br m, 1H), 1.71 (br d, J=13 Hz, 1H).

Step 9. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(hydroxymethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (P1)

Boron trichloride (1 M solution in heptane, 89.7 mL, 89.7 mmol) was added to a 0° C. solution of N-[(4aR,6R,8aS)-6-[(benzyloxy)methyl]-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C8) (15.20 g, 29.89 mmol) in dichloromethane (150 mL). After 15 minutes, the reaction mixture was allowed to warm to room temperature and stirred for 4 hours. Methanol (50 mL) was then added, first drop-wise {Caution: violent reaction} and then at a steady rate, while the interior of the flask was flushed with nitrogen gas. The mixture was heated at reflux for 30 minutes, cooled to room temperature and concentrated in vacuo. The residue was again dissolved in methanol, stirred, and concentrated in vacuo. The resulting material was taken up in dichloromethane and washed sequentially with 1 M aqueous sodium hydroxide solution, water, and saturated aqueous sodium chloride solution. The organic layer was dried over magnesium sulfate, filtered, and concentrated under reduced pressure. Chromatographic purification on silica gel (Gradient: 0% to 3% methanol in ethyl acetate) provided the product as a yellow foam. Yield: 11.97 g, 28.60 mmol, 96%. LCMS m/z 419.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 8.13 (d, J=7.4 Hz, 2H), 7.50-7.56 (m, 1H), 7.41-7.49 (m, 3H), 7.02-7.11 (m, 2H), 4.13 (dd, J=11.9, 1.8 Hz, 1H), 3.90 (d, J=12.1 Hz, 1H), 3.72-3.80 (m, 1H), 3.59 (d, J=5.1 Hz, 2H), 3.14-3.24 (br m, 1H), 2.96 (dd, half of ABX pattern, J=13.1, 4.1 Hz, 1H), 2.75 (dd, half of ABX pattern, J=13.1, 2.7 Hz, 1H), 1.80-1.92 (m, 1H), 1.70 (ddd, J=13.4, 4.2, 2.4 Hz, 1H).

Alternate conversion of ({[(2R)-2-(2,2-diethoxyethoxy)pent-4-en-1-yl]oxy}methyl)benzene (C2) to (3aR,5R)-5-[(benzyloxy)methyl]-3,3a,4,5-tetrahydro-7H-pyrano[3,4-c][1,2]oxazole (C4)

Step 1. Synthesis of 2-{[(2R)-1-(benzyloxy)pent-4-en-2-yl]oxy}-N-hydroxyethanimine (C3)

({[(2R)-2-(2,2-Diethoxyethoxy)pent-4-en-1-yl]oxy}methyl)benzene (C2) (12.4 g, 40.2 mmol) was dissolved in acetic acid (28 mL) and water (12 mL). Hydroxylamine hydrochloride (2.84 g, 40.9 mmol) was added as a solid. After 1 hour, additional hydroxylamine hydrochloride (2.84 g, 40.9 mmol) was added. After 1 more hour, the reaction mixture was diluted with tert-butyl methyl ether (100 mL) and washed with water (3×50 mL), then washed with aqueous potassium carbonate solution (0.5 M, 100 mL). The organic layer was concentrated to provide the product as a pale yellow oil, which consisted of a roughly equimolar mixture of oxime isomers, as assessed by ¹H NMR. Yield: 9.60 g, 38.5 mmol, 96%. ¹H NMR (400 MHz, CDCl₃) δ 7.98 and 7.67 (2 br s, total 1H), [7.50 (t, J=5.6 Hz) and 6.95 (t, J=3.6 Hz), total 1H], 7.28-7.39 (m, 5H), 5.74-5.87 (m, 1H), 5.04-5.14 (m, 2H), 4.55 and 4.56 (2 s, total 2H), 4.47-4.49 (m, 1H), 4.18-4.28 (m, 1H), 3.47-3.65 (m, 3H), 2.30-2.37 (m, 2H).

Step 2. Synthesis of (3aR,5R)-5-[(benzyloxy)methyl]-3,3a,4,5-tetrahydro-7H-pyrano[3,4-c][1,2]oxazole (C4)

Pyridine (23.1 mL, 286 mmol) was added to a solution of 2-{[(2R)-1-(benzyloxy)pent-4-en-2-yl]oxy}-N-hydroxyethanimine (C3) (35.6 g, 143 mmol) in dichloromethane (350 mL). N-Chlorosuccinimide (19.4 g, 145 mmol) was added in portions over roughly 2 hours. The reaction was stirred for 3 hours, then diluted with an aqueous solution of sodium sulfite (5 g in 100 mL water). The mixture was stirred for 20 minutes, and the aqueous layer was extracted with dichloromethane; the combined organic layers were washed with water, dried, and concentrated. Purification via silica gel chromatography (Eluent: 1:2 ethyl acetate/hexanes) afforded the product. Yield: 21.2 g, 85.7 mmol, 60%. ¹H NMR (400 MHz, CDCl₃) δ 7.28-7.40 (m, 5H), 4.77 (d, J=13.4 Hz, 1H), 4.55-4.65 (m, 3H), 4.22 (dd, J=13.5, 1.3 Hz, 1H), 3.79 (dd, J=11.7, 8.0 Hz, 1H), 3.69-3.76 (m, 1H), 3.57 (dd, half of ABX pattern, J=10.2, 5.9 Hz, 1H), 3.49 (dd, half of ABX pattern, J=10.2, 4.3 Hz, 1H), 3.40-3.5 (m, 1H), 2.21 (ddd, J=12.9, 6.5, 1.8 Hz, 1H), 1.57 (ddd, J=13, 12, 11 Hz, 1H).

Preparation P2 N-[(4R,4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(hydroxymethyl)-4-methyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (P2)

Step 1. Synthesis of (2R)-1-(benzyloxy)hex-4-en-2-ol (C9)

The product was obtained according to the method used for synthesis of (2R)-1-(benzyloxy)pent-4-en-2-ol (C1) in Preparation P1, except that 1-propenylmagnesium bromide was used in place of vinylmagnesium bromide. The product was obtained as a brown oil, which was used without further purification; by ¹H NMR, this material consisted of a 1:1 mixture of geometric isomers. Yield: 140 g, 0.679 mol, 100%. ¹H NMR (400 MHz, CDCl₃) δ 7.28-7.42 (m, 5H), 5.39-5.67 (m, 2H), 4.57 (s, 2H), 3.80-3.92 (m, 1H), 3.48-3.57 (m, 1H), 3.35-3.43 (m, 1H), 2.36-2.50 (br m, 1H), 2.24-2.33 (m, 1H), 2.17-2.24 (m, 1H), [1.68 (br d, J=6 Hz) and 1.64 (br d, J=7 Hz), total 3H].

Step 2. Synthesis of ({[(2R)-2-(2,2-diethoxyethoxy)hex-4-en-1-yl]oxy}methyl)benzene (C10)

(2R)-1-(Benzyloxy)hex-4-en-2-ol (C9) (150 g, 0.73 mol) was converted to the product according to the method used for synthesis of ({[(2R)-2-(2,2-diethoxyethoxy)pent-4-en-1-yl]oxy}methyl)benzene (C2) in Preparation P1, except that the initial combination of reagents was carried out at 0° C. The product was obtained as a brown oil (400 g, ≦0.73 mol), which was used for the next step without further purification. By ¹H NMR analysis, this material contained a roughly 1:1 mixture of geometric isomers. ¹H NMR (400 MHz, CDCl₃), characteristic peaks for product: δ 7.25-7.38 (m, 5H), 5.38-5.60 (m, 2H), 4.55 and 4.55 (2 s, total 2H), 2.22-2.37 (m, 2H), 1.60-1.68 (m, 3H).

Step 3. Synthesis of {[(2R)-1-(benzyloxy)hex-4-en-2-yl]oxy}acetaldehyde (C11)

To a solution of ({[(2R)-2-(2,2-diethoxyethoxy)hex-4-en-1-yl]oxy}methyl)benzene (C10) (350 g from the previous step, ≦0.64 mol) in tetrahydrofuran (1.4 L) was added aqueous hydrochloric acid (2 M, 700 mL), and the reaction mixture was stirred at 75° C. for 1 hour. Solvent was removed in vacuo and the aqueous residue was extracted with ethyl acetate (2.0 L). The combined organic layers were washed with saturated aqueous sodium chloride solution (3×500 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. The product was obtained as a pale brown oil (210 g, ≦0.64 mol), which was taken directly to the following step.

Step 4. Synthesis of 2-{[(2R)-1-(benzyloxy)hex-4-en-2-yl]oxy}-N-hydroxyethanimine (C12)

To a mixture of {[(2R)-1-(benzyloxy)hex-4-en-2-yl]oxy}acetaldehyde (C11) (207 g, ≦0.63 mol) and sodium acetate (342 g, 4.17 mol) in aqueous ethanol (2:1 ethanol/water, 2.1 L) was added hydroxylamine hydrochloride (207 g, 2.98 mol). The reaction mixture was stirred at 60° C. for 18 hours, then concentrated in vacuo and extracted with ethyl acetate (2.0 L). The combined organic layers were dried over sodium sulfate, filtered, concentrated under reduced pressure and purified by chromatography on silica gel (Eluent: ethyl acetate in petroleum ether) to afford the product as a brown oil. By ¹H NMR, this was assigned as a mixture of geometric isomers at both the oxime and olefin functional groups. Yield: 117 g, 0.444 mol, 70% over three steps. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ [7.42-7.48 (m) and 6.88-6.92 (m), total 1H], 7.20-7.36 (m, 5H), 5.29-5.61 (m, 2H), [4.48-4.54 (m) and 4.41-4.45 (m), total 3H], 2.13-2.32 (m, 2H), 1.54-1.65 (m, 3H).

Step 5. Synthesis of (3S,3aR,5R)-5-[(benzyloxy)methyl]-3-methyl-3,3a,4,5-tetrahydro-7H-pyrano[3,4-c][1,2]oxazole (C13) and (3R,3aR,5R)-5-[(benzyloxy)methyl]-3-methyl-3,3a,4,5-tetrahydro-7H-pyrano[3,4-c][1,2]oxazole (C14)

An aqueous solution of sodium hypochlorite (6.15% solution, 6.6 L) was slowly added to a solution of 2-{[(2R)-1-(benzyloxy)hex-4-en-2-yl]oxy}-N-hydroxyethanimine (C12) (660 g, 2.51 mol) and triethylamine (19 g, 0.19 mol) in dichloromethane (6.6 L) at 25° C. After completion of the addition, the reaction mixture was stirred at 25° C. for 30 minutes. The organic layer was washed with water (3×3 L), dried over sodium sulfate, filtered, and concentrated in vacuo. Purification via chromatography on silica gel (Eluent: ethyl acetate in petroleum ether) provided (3S,3aR,5R)-5-[(benzyloxy)methyl]-3-methyl-3,3a,4,5-tetrahydro-7H-pyrano[3,4-c][1,2]oxazole (C13) as a white solid. Yield: 90 g, 0.34 mol, 14%. The indicated relative stereochemistry of compound C13 was assigned based on nuclear Overhauser enhancement studies, which revealed interactions of the methine proton on carbon 3a with both the protons of the methyl group on carbon 3 and the methine proton on carbon 5. LCMS m/z 261.9 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.24-7.39 (m, 5H), 4.69 (d, J=13.7 Hz, 1H), 4.57 (AB quartet, J_(AB)=12.2 Hz, Δν_(AB)=13.8 Hz, 2H), 4.13-4.25 (m, 2H), 3.62-3.70 (m, 1H), 3.55 (dd, half of ABX pattern, J=10, 6 Hz, 1H), 3.47 (dd, half of ABX pattern, J=10, 4 Hz, 1H), 2.93 (br ddd, J=11, 11, 7 Hz, 1H), 2.11 (br dd, J=12.6, 6.8 Hz, 1H), 1.45-1.56 (m, 1H), 1.45 (d, J=6.2 Hz, 3H).

Also obtained from the chromatographic separation was (3R,3aR,5R)-5-[(benzyloxy)methyl]-3-methyl-3,3a,4,5-tetrahydro-7H-pyrano[3,4-c][1,2]oxazole (C14), as a brown oil. Yield: 126 g, 0.482 mol, 19%. The indicated relative stereochemistry of compound C14 was assigned based on nuclear Overhauser enhancement studies, which revealed interactions of the methine proton on carbon 3a with both the methine proton on carbon 3 and the methine proton on carbon 5. LCMS m/z 261.9 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.26-7.39 (m, 5H), 4.76-4.86 (m, 1H), 4.75 (d, J=13.5 Hz, 1H), 4.58 (AB quartet, J_(AB)=12.2 Hz, Δν_(AB)=12.4 Hz, 2H), 4.19 (dd, J=13.5, 1.2 Hz, 1H), 3.63-3.70 (m, 1H), 3.57 (dd, half of ABX pattern, J=10.2, 6.0 Hz, 1H), 3.49 (dd, half of ABX pattern, J=10.1, 4.2 Hz, 1H), 3.36 (br ddd, J=11.4, 11.4, 6.3 Hz, 1H), 1.86 (ddd, J=12.8, 6.4, 1.2 Hz, 1H), 1.55-1.66 (m, 1H), 1.16 (d, J=6.6 Hz, 3H).

Step 6. Synthesis of (3S,3aR,5R,7aS)-5-[(benzyloxy)methyl]-7a-(2,4-difluorophenyl)-3-methylhexahydro-1H-pyrano[3,4-c][1,2]oxazole (C15)

The product, obtained as a yellow oil, was prepared from (3S,3aR,5R)-5-[(benzyloxy)methyl]-3-methyl-3,3a,4,5-tetrahydro-7H-pyrano[3,4-c][1,2]oxazole (C13) according to the general procedure for the synthesis of (3aR,5R,7aS)-5-[(benzyloxy)methyl]-7a-(2,4-difluorophenyl)hexahydro-1H-pyrano[3,4-c][1,2]oxazole (C5) in Preparation P1. Yield: 21.5 g, 57.2 mmol, 48%. LCMS m/z 376.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.98 (ddd, J=9.1, 9.1, 6.8 Hz, 1H), 7.28-7.40 (m, 5H), 6.87-6.93 (m, 1H), 6.80 (ddd, J=11.9, 8.6, 2.6 Hz, 1H), 4.60 (AB quartet, J_(AB)=12.1 Hz, Δν_(AB)=19.9 Hz, 2H), 3.99-4.06 (m, 1H), 3.97 (dd, half of ABX pattern, J=12.9, 2.0 Hz, 1H), 3.80-3.88 (m, 2H), 3.56 (dd, half of ABX pattern, J=10.2, 6.3 Hz, 1H), 3.49 (dd, half of ABX pattern, J=10.2, 4.1 Hz, 1H), 2.81-2.87 (m, 1H), 2.04 (ddd, J=14.2, 7.6, 2.8 Hz, 1H), 1.48-1.59 (m, 1H), 0.79 (d, J=6.4 Hz, 3H).

Step 7. Synthesis of (1S)-1-[(2R,4R,5S)-5-amino-2-[(benzyloxy)methyl]-5-(2,4-difluorophenyl)tetrahydro-2H-pyran-4-yl]ethanol (C16)

The product, obtained as a yellow oil, was prepared from (3S,3aR,5R,7aS)-5-[(benzyloxy)methyl]-7a-(2,4-difluorophenyl)-3-methylhexahydro-1H-pyrano[3,4-c][1,2]oxazole (C15) according to the general procedure for the synthesis of [(2R,4R,5S)-5-amino-2-[(benzyloxy)methyl]-5-(2,4-difluorophenyl)tetrahydro-2H-pyran-4-yl]methanol (C6) in Preparation P1. Yield: 13.96 g, 37.00 mmol, 98%. LCMS m/z 378.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ 7.65-7.78 (br m, 1H), 7.27-7.40 (m, 5H), 6.93-7.02 (br m, 1H), 6.80 (ddd, J=12.6, 8.5, 2.6 Hz, 1H), 4.06 (dd, J=11.7, 2.2 Hz, 1H), 3.53 (dd, J=10.2, 3.7 Hz, 1H), 2.50-2.61 (br m, 1H), 1.62 (ddd, J=14, 4, 2.5 Hz, 1H), 0.89 (d, J=6.6 Hz, 3H).

Step 8. Synthesis of N-{[(3S,4R,6R)-6-[(benzyloxy)methyl]-3-(2,4-difluorophenyl)-4-[(1S)-1-hydroxyethyl]tetrahydro-2H-pyran-3-yl]carbamothioyl}benzamide (C17)

The product was prepared from (1S)-1-[(2R,4R,5S)-5-amino-2-[(benzyloxy)methyl]-5-(2,4-difluorophenyl)tetrahydro-2H-pyran-4-yl]ethanol (C16) according to the general procedure for the synthesis of N-{[(3S,4R,6R)-6-[(benzyloxy)methyl]-3-(2,4-difluorophenyl)-4-(hydroxymethyl)tetrahydro-2H-pyran-3-yl]carbamothioyl}benzamide (C7) in Preparation P1. In this case, after concentration of the reaction mixture in vacuo, the residue was chromatographed on silica gel (Gradient: 0% to 50% ethyl acetate in heptane) to afford the product as a yellow foam. Yield: 13.36 g, 24.71 mmol, 67%. LCMS m/z 539.2 [M−H⁺].

Step 9. Synthesis of N-[(4R,4aR,6R,8aS)-6-[(benzyloxy)methyl]-8a-(2,4-difluorophenyl)-4-methyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C18)

Diethyl azodicarboxylate (21.3 mL, 136 mmol) was added drop-wise to a solution of triphenylphosphine (35.7 g, 136 mmol) in tetrahydrofuran (850 mL), and the mixture was stirred for 30 minutes before being cooled in an ice bath. A solution of N-{[(3S,4R,6R)-6-[(benzyloxy)methyl]-3-(2,4-difluorophenyl)-4-[(1S)-1-hydroxyethyl]tetrahydro-2H-pyran-3-yl]carbamothioyl}benzamide (C17) (24.5 g, 45.3 mmol) in tetrahydrofuran (115 mL) was added drop-wise to the reaction mixture, which was then stirred for 1 hour under ice cooling. After concentration in vacuo, the residue was loaded onto a silica gel column that had been equilibrated with dichloromethane, and the column was eluted with 1:1 ethyl acetate/heptane. Fractions containing product were combined and concentrated under reduced pressure; the resulting material was triturated with 15% ethyl acetate in heptane, and the solid was removed via filtration. The filtrate was concentrated in vacuo and chromatographed on silica gel (Gradient: 20% to 40% ethyl acetate in heptane), affording the product as a white solid. Yield: 17.23 g, 32.97 mmol, 73%. LCMS m/z 523.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.23 (br d, J=6.5 Hz, 2H), 7.49-7.55 (m, 1H), 7.36-7.48 (m, 3H), 7.24-7.36 (m, 5H), 6.84-6.96 (m, 2H), 4.58 (AB quartet, J_(AB)=12.0 Hz, Δν_(AB)=25.0 Hz, 2H), 4.18 (dd, J=12.2, 1.7 Hz, 1H), 3.87-3.94 (m, 1H), 3.84 (d, J=12.2 Hz, 1H), 3.63 (dd, half of ABX pattern, J=10.2, 6.4 Hz, 1H), 3.50 (dd, half of ABX pattern, J=10.2, 4.4 Hz, 1H), 3.23-3.31 (m, 1H), 2.88-2.96 (m, 1H), 1.61-1.79 (m, 2H), 1.25 (d, J=6.9 Hz, 3H).

Step 10. Synthesis of N-[(4R,4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(hydroxymethyl)-4-methyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (P2)

The product was prepared from N-[(4R,4aR,6R,8aS)-6-[(benzyloxy)methyl]-8a-(2,4-difluorophenyl)-4-methyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C18) according to the general procedure for the synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(hydroxymethyl)-4,4a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (P1) in Preparation P1. In this case, the combined crude product from two similar reactions was triturated with dichloromethane rather than being purified by chromatography. The filtrate from the trituration was concentrated in vacuo, and a second crop of material was obtained via a second trituration with dichloromethane, affording the product in both cases as a white solid. Total yield: 23.12 g, 53.46 mmol, 79%. LCMS m/z 433.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 8.12 (br d, J=7 Hz, 2H), 7.51-7.57 (m, 1H), 7.40-7.49 (m, 3H), 7.02-7.11 (m, 2H), 4.15 (br d, J=12 Hz, 1H), 3.91 (d, J=11.9 Hz, 1H), 3.71-3.78 (m, 1H), 3.60 (d, J=5.2 Hz, 2H), 3.19-3.28 (br m, 1H), 2.97-3.06 (br m, 1H), 1.74-1.82 (m, 1H), 1.49-1.62 (m, 1H), 1.26 (d, J=7.0 Hz, 3H).

Example 1 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (1)

Step 1. Synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazine-6-carboxylic acid (C19)

Tetrapropylammonium perruthenate (209 mg, 0.595 mmol) was added to a mixture of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(hydroxymethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (P1) (2.49 g, 5.95 mmol) and 4-methylmorpholine N-oxide monohydrate (4.83 g, 35.7 mmol) in acetonitrile (125 mL), and the reaction mixture was stirred for 40 minutes at room temperature. After addition of 2-propanol (50 mL), it was stirred for an additional 2 hours and then concentrated in vacuo. The residue was partitioned between 1:1 ethyl acetate/diethyl ether (150 mL) and aqueous sodium hydroxide solution (0.25 M, 150 mL). The organic layer was extracted with aqueous sodium hydroxide solution (0.25 M, 2×150 mL), and the combined aqueous layers were acidified to a pH of approximately 1 with 2 M aqueous hydrochloric acid, then extracted with ethyl acetate (3×200 mL). The combined ethyl acetate layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. Purification via chromatography on silica gel [Gradient: 0% to 100% (89:10:1 dichloromethane/methanol/acetic acid) in dichloromethane] provided the product as a purple-white solid. Yield: 2.40 g, 5.55 mmol, 93%. LCMS m/z 433.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 8.08-8.13 (m, 2H), 7.52-7.57 (m, 1H), 7.43-7.50 (m, 3H), 7.02-7.11 (m, 2H), 4.36 (dd, J=11.2, 3.4 Hz, 1H), 4.19 (br d, J=12 Hz, 1H), 3.97 (d, J=12.1 Hz, 1H), 3.20-3.27 (m, 1H), 2.96 (dd, half of ABX pattern, J=13.2, 4.0 Hz, 1H), 2.78 (dd, half of ABX pattern, J=13.2, 2.8 Hz, 1H), 2.02-2.15 (m, 2H).

Step 2. Synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C20)

A mixture of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylic acid (C19) [1.26 g, 2.91 mmol; this material had been azeotroped with toluene (2×5 mL)], N,N-diisopropylethylamine (1.02 mL, 5.86 mmol) and 2-[2-oxo-1(2H)-pyridyl]-1,1,3,3-tetramethyluronium tetrafluoroborate (TPTU, 866 mg, 2.91 mmol) in N,N-dimethylformamide (10 mL) was stirred at room temperature for 25 minutes. 2,2-Dimethoxyethanamine (0.95 mL, 8.7 mmol) was added, and stirring was continued for 18 hours, at which time the reaction mixture was diluted with saturated aqueous sodium bicarbonate solution (15 mL) and water (15 mL), and extracted with diethyl ether (3×40 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting oil was combined with material derived from a similar reaction carried out on C19 (131 mg, 0.303 mmol) and purified via silica gel chromatography (Gradient: 0% to 100% ethyl acetate in heptane) to afford the product as a white solid. Yield: 618 mg, 1.19 mmol, 37%. LCMS m/z 520.3 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.17-8.28 (br m, 2H), 7.50-7.56 (m, 1H), 7.36-7.49 (m, 3H), 6.86-6.99 (m, 2H), 6.74 (br t, J=6 Hz, 1H), 4.38 (t, J=5.3 Hz, 1H), 4.15-4.24 (m, 2H), 3.90 (d, J=12.3 Hz, 1H), 3.48-3.57 (m, 1H), 3.37 (s, 3H), 3.36 (s, 3H), 3.28-3.36 (m, 1H), 3.13-3.23 (br m, 1H), 3.03 (dd, J=12.9, 3.9 Hz, 1H), 2.70 (br d, J=13 Hz, 1H), 2.20-2.29 (m, 1H), 2.00-2.13 (m, 1H).

Step 3. Synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2-oxoethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C21)

(4aR,6R,8aS)-2-(Benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C20) (620 mg, 1.19 mmol) was dissolved in tetrahydrofuran (3.0 mL) and aqueous hydrochloric acid (2 M, 3 mL, 6 mmol), and stirred for 18 hours at 38° C. After cooling to room temperature, the reaction mixture was diluted with saturated aqueous sodium chloride solution (7 mL) and extracted with ethyl acetate (6×10 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. Purification via silica gel chromatography (Gradient: 50% to 100% ethyl acetate in heptane) provided the product as an off-white solid. Yield: 352 mg, 0.743 mmol, 62%. LCMS m/z 474.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 9.63 (s, 1H), 8.34-8.44 (br m, 2H), 7.64-7.71 (br m, 1H), 7.54-7.62 (br m, 2H), 7.40-7.51 (br m, 1H), 7.28-7.37 (br m, 1H), 7.00-7.10 (br m, 1H), 6.91-7.00 (br m, 1H), 4.36-4.50 (br m, 1H), 4.23-4.35 (br m, 2H), 4.19 (br d, J=11 Hz, 1H), 4.00-4.11 (br m, 1H), 3.29-3.46 (br m, 1H), 3.01-3.13 (br m, 1H), 2.76-2.89 (br m, 1H), 2.28-2.44 (br m, 1H), 1.85-2.02 (br m, 1H).

Step 4. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C22)

A mixture of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2-oxoethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazine-6-carboxamide (C21) [38 mg, 80 μmol; this material had been azeotroped with toluene (2×2 mL)] and Burgess reagent (1-methoxy-N-triethylammoniosulfonylmethanimidate, 38.1 mg, 0.160 mmol) in tetrahydrofuran (2.5 mL) was heated at reflux for 1.5 hours, at which time it was cooled to room temperature and treated with additional Burgess reagent (19 mg, 80 μmol). After an additional 3 hours of heating at reflux, the reaction mixture was again allowed to cool to room temperature, then concentrated in vacuo. Purification via silica gel chromatography (0% to 100% ethyl acetate in heptane) afforded the product as a white solid. Yield: 12 mg, 26 μmol, 32%. LCMS m/z 456.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 8.11 (br d, J=7.4 Hz, 2H), 7.92-7.93 (m, 1H), 7.43-7.58 (m, 4H), 7.17-7.18 (m, 1H), 7.04-7.14 (m, 2H), 4.97 (dd, J=12, 2 Hz, 1H), 4.32 (br d, J=12 Hz, 1H), 4.01 (d, J=11.9 Hz, 1H), 3.3-3.37 (m, 1H, assumed; obscured by solvent peak), 3.00 (dd, half of ABX pattern, J=13.1, 4.1 Hz, 1H), 2.82 (dd, half of ABX pattern, J=13, 2.6 Hz, 1H), 2.40-2.52 (m, 1H), 2.04-2.11 (m, 1H).

Step 5. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-amine (1)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C22) (25.0 mg, 54.9 μmol) was combined with methanol (2.0 mL) and 1,8-diazabicyclo[5.4.0]undec-7-ene (2 drops) and heated to 75° C. for 18 hours. The reaction mixture was cooled and concentrated in vacuo; purification using silica gel chromatography (Gradient: 0% to 10% methanol in dichloromethane) provided the product as a white solid. Yield: 11.2 mg, 31.9 μmol, 58%. LCMS m/z 352.1 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 7.92 (d, J=0.8 Hz, 1H), 7.38 (ddd, J=9.6, 8.8, 6.6 Hz, 1H), 7.17 (d, J=0.8 Hz, 1H), 6.95-7.04 (m, 2H), 4.87 (dd, J=11.9, 2.3 Hz, 1H), 4.27 (dd, J=11.2, 2.0 Hz, 1H), 3.80 (d, J=11.3 Hz, 1H), 3.03-3.10 (m, 1H), 2.93 (dd, half of ABX pattern, J=12.6, 4.2 Hz, 1H), 2.76 (dd, half of ABX pattern, J=12.6, 2.8 Hz, 1H), 2.35-2.46 (m, 1H), 1.90 (ddd, J=13.3, 4.1, 2.5 Hz, 1H).

Example 2 and Example 3 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1-methyl-1H-pyrazol-3-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (2) and (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1-methyl-1H-pyrazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-amine (3)

Step 1. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-formyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C23)

Triethylamine (16.7 mL, 120 mmol) was added in one rapid portion to a solution of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(hydroxymethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (P1) (4.18 g, 10.0 mmol) in dichloromethane (200 mL) that was immersed in a room temperature water bath. After 5 minutes, anhydrous dimethyl sulfoxide (9.94 mL, 140 mmol) was rapidly added, followed immediately by solid sulfur trioxide pyridine complex (98%, 13.0 g, 80.0 mmol) in a single portion. The resulting solution was stirred at ambient temperature for 6.5 hours, then diluted with a 1:1 mixture of water and saturated aqueous sodium chloride solution (200 mL) and stirred for 10 minutes. The aqueous layer was extracted with dichloromethane (2×200 mL), and the combined organic layers were washed with water (100 mL), washed with saturated aqueous sodium chloride solution (100 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Purification via silica gel chromatography (Gradient: 0% to 100% ethyl acetate in heptane) gave the product as a white solid. Yield: 2.81 g, 6.75 mmol, 67%. LCMS m/z 414.9 [M−H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 9.71 (s, 1H), 8.20 (br d, J=7 Hz, 2H), 7.50-7.56 (m, 1H), 7.36-7.49 (m, 3H), 6.86-6.99 (m, 2H), 4.23 (br d, J=12.1 Hz, 1H), 4.12 (dd, J=12.1, 2.9 Hz, 1H), 3.94 (d, J=12.5 Hz, 1H), 3.13-3.22 (m, 1H), 3.04 (dd, J=13.1, 4.1 Hz, 1H), 2.69 (dd, J=13.1, 2.9 Hz, 1H), 2.02-2.14 (m, 1H), 1.92-1.99 (m, 1H).

Step 2. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-hydroxyprop-2-yn-1-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C24)

A solution of ethynylmagnesium bromide in tetrahydrofuran (0.5 M, 8.0 mL, 4.0 mmol) was cooled to 15° C. A solution of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-formyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C23) (416 mg, 1.00 mmol) in tetrahydrofuran (10 mL) was then added drop-wise over 15 minutes, during which time the internal reaction temperature rose to 23° C. The reaction mixture was stirred at room temperature for an additional 30 minutes, then treated with additional Grignard reagent (1 mL, 0.5 mmol). As no change was detected by thin layer chromatography, the reaction mixture was cooled to 0° C., quenched with saturated aqueous ammonium chloride solution (15 mL) and extracted with ethyl acetate (3×20 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to afford the crude product as an amber foam (466 mg, assumed quantitative), which was taken directly to the following step. LCMS m/z 443.2 [M+H⁺].

Step 3. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(prop-2-ynoyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C25)

Dess-Martin periodinane [1,1,1-tris(acetyloxy)-1,1-dihydro-1,2-benziodoxol-3-(1H)-one] (458 mg, 1.08 mmol) was added to a 0° C. solution of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-hydroxyprop-2-yn-1-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C24) (material from the previous step, 466 mg, ≦1.0 mmol) in dichloromethane (12 mL). The reaction mixture was allowed to warm to room temperature and then stirred for 1.5 hours. After addition of dichloromethane (40 mL), saturated aqueous sodium thiosulfate solution (20 mL) and saturated aqueous sodium bicarbonate solution (30 mL) were added and the mixture was stirred for 30 minutes. The organic layer was washed with saturated aqueous sodium bicarbonate solution (20 mL) and with saturated aqueous sodium chloride solution (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Purification using silica gel chromatography (Gradient: 0% to 65% ethyl acetate in heptane) afforded the product as an off-white solid. Yield: 291 mg, 0.66 mmol, 66% over two steps. LCMS m/z 441.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.18 (br d, J=7.6 Hz, 2H), 7.50-7.57 (m, 1H), 7.36-7.49 (m, 3H), 6.86-6.99 (m, 2H), 4.28 (dd, J=11.5, 3.1 Hz, 1H), 4.22 (dd, J=12.2, 1.5 Hz, 1H), 3.97 (d, J=12.1 Hz, 1H), 3.42 (s, 1H), 3.15-3.25 (m, 1H), 3.04 (dd, J=13.0, 4.0 Hz, 1H), 2.69 (dd, J=13.1, 2.7 Hz, 1H), 2.07-2.25 (m, 2H).

Step 4. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-3-yl)-4,4a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C26) and N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C27)

A slurry of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(prop-2-ynoyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C25) (55 mg, 0.12 mmol) and methylhydrazine (6.60 μL, 0.125 mmol) in 2-propanol (2 mL) was stirred at room temperature for 3 hours, then concentrated under a stream of nitrogen to afford an off-white foam (52 mg). This was combined with material (20 mg) from an identical reaction carried out on C25 (19.8 mg, 45 μmol) and purified via silica gel chromatography (Gradient: 25% to 100% ethyl acetate in heptane) to afford the product as an off-white foam, determined to be a roughly 4:1 mixture of C26 and C27 by ¹H NMR analysis. Yield: 50.5 mg, 0.108 mmol, 64%. LCMS m/z 469.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), peaks for C26 only: δ 8.22 (br d, J=7.4 Hz, 2H), 7.40-7.55 (m, 4H), 7.29 (d, J=2.2 Hz, 1H), 6.86-6.97 (m, 2H), 6.29 (d, J=2.2 Hz, 1H), 4.83 (dd, J=11.6, 2.3 Hz, 1H), 4.33 (dd, J=12.2, 1.5 Hz, 1H), 3.9-3.96 (m, 1H), 3.87 (s, 3H), 3.24-3.33 (m, 1H), 3.06 (dd, J=12.9, 4.1 Hz, 1H), 2.69 (dd, J=12.9, 2.7 Hz, 1H), 2.34-2.47 (m, 1H), 1.99-2.07 (m, 1H). Characteristic peaks for minor isomer C27 that were observed in this spectrum: 8.16-8.20 (m, 1H), 6.24 (br d, J=1.6 Hz, 1H), 4.79 (dd, J=11.9, 2.2 Hz, 1H), 3.93 (s, 3H).

Step 5. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-3-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (2) and (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (3)

A mixture of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-3-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C26) and N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C27) (−4:1 mixture from the preceding step, 50.5 mg, 0.108 mmol) was dissolved in methanol (0.5 mL) and treated with 1,8-diazabicyclo[5.4.0]undec-7-ene (13 μL, 87 μmol). The reaction mixture was heated at 80° C. for 18 hours, then concentrated under a stream of nitrogen and partitioned between water (2 mL) and ethyl acetate (6 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated in vacuo. After purification via silica gel chromatography (Gradient: 0% to 15% methanol in dichloromethane), (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-3-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (2) was obtained as an off-white solid. Yield: 23 mg, 63 μmol, 58%. LCMS m/z 365.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.44 (ddd, J=9.1, 9.0, 6.7 Hz, 1H), 7.29 (d, J=2.2 Hz, 1H), 6.84-6.90 (m, 1H), 6.80 (ddd, J=12.4, 8.6, 2.6 Hz, 1H), 6.28 (d, J=2.2 Hz, 1H), 4.76 (dd, J=11.7, 2.2 Hz, 1H), 4.25 (dd, J=11.2, 2.3 Hz, 1H), 3.96 (d, J=11.3 Hz, 1H), 3.90 (s, 3H), 2.96-3.08 (m, 2H), 2.65 (dd, J=12.0, 2.4 Hz, 1H), 2.21-2.32 (m, 1H), 1.81 (ddd, J=13.4, 3.8, 2.4 Hz, 1H).

Fractions containing (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (3) were subjected to purification via reversed-phase HPLC (Column: Waters XBridge C18, 5 μm; Mobile phase A: 0.03% ammonium hydroxide in water (v/v); Mobile phase B: 0.03% ammonium hydroxide in acetonitrile (v/v); Gradient: 20% to 60% B) to provide 3 as a solid. Yield: 6 mg, 16 μmol, 15%. LCMS m/z 365.0 [M+H⁺]. ¹H NMR (600 MHz, DMSO-d₆), observed peaks: δ 7.39-7.45 (m, 1H), 7.36 (d, J=1.8 Hz, 1H), 7.32-7.37 (m, 1H), 7.24-7.29 (m, 1H), 6.28 (d, J=1.8 Hz, 1H), 4.98 (dd, J=11.2, 2.0 Hz, 1H), 4.20 (d, J=11.8 Hz, 1H), 3.96 (d, J=12.3 Hz, 1H), 3.85 (s, 3H), 3.10 (dd, J=13.2, 2.6 Hz, 1H), 2.95 (dd, J=12.9, 4.2 Hz, 1H), 2.05-2.10 (m, 1H), 1.94-2.02 (m, 1H). For both products, the pyrazole regiochemistry was assigned via nuclear Overhauser enhancement studies.

Example 4 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(4-methyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (4)

Step 1. Synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(1-hydroxypropan-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C28)

The product was synthesized according to the general procedure for the synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C20) in Example 1, by utilizing 2-aminopropan-1-ol in place of 2,2-dimethoxyethanamine. In this case, at the completion of the reaction, the reaction mixture was diluted with saturated aqueous sodium bicarbonate solution and extracted three times with tert-butyl methyl ether. The combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo. Purification using silica gel chromatography (Gradient: 0% to 100% ethyl acetate in heptane, followed by isocratic elution with 20% methanol in dichloromethane), provided the product as a yellow foam. ¹H NMR analysis revealed this material to consist of a 1:1 mixture of diastereomers, in a 1.3:1 ratio with N,N-dimethylformamide. Corrected yield: 2.56 g, 5.24 mmol, 87%. LCMS m/z 488.1 [M−H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 8.11 (br d, J=7 Hz, 2H), 7.43-7.58 (m, 4H), 7.03-7.13 (m, 2H), 4.17-4.25 (m, 2H), 3.95-4.05 (m, 2H), 3.48-3.53 (m, 2H), 3.18-3.28 (br m, 1H), 2.97 (dd, J=13.2, 4.1 Hz, 1H), 2.77-2.83 (m, 1H), 2.09-2.17 (br m, 1H), 1.86-2.02 (m, 1H), [1.16 (d, J=6.8 Hz) and 1.13 (d, J=6.8 Hz), total 3H].

Step 2. Synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(1-oxopropan-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C29)

The product was prepared from (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(1-hydroxypropan-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazine-6-carboxamide (C28) according to the procedure for preparation of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(prop-2-ynoyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C25) in Example 2/Example 3, and was obtained as a pale yellow foam. ¹H NMR indicated this to consist of a roughly 1:1 mixture of diastereomers. Yield: 2.11 g, 4.33 mmol, 83%. ¹H NMR (400 MHz, CDCl₃) 6 [9.56 (d, J=0.2 Hz) and 9.53 (d, J=0.4 Hz), total 1H], 8.18-8.25 (m, 2H), 7.51-7.56 (m, 1H), 7.36-7.49 (m, 3H), 7.02-7.10 (m, 1H), 6.87-6.99 (m, 2H), 4.37-4.54 (m, 1H), 4.19-4.27 (m, 2H), 3.93 (d, J=12.3 Hz, 1H), 3.15-3.24 (br m, 1H), 3.00-3.07 (m, 1H), 2.68-2.74 (m, 1H), 2.22-2.29 (m, 1H), 2.04-2.17 (m, 1H), [1.39 (d, J=7.3 Hz) and 1.39 (d, J=7.5 Hz), total 3H].

Step 3. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(4-methyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C30)

A mixture of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(1-oxopropan-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazine-6-carboxamide (C29) (2.11 g, 4.33 mmol) and Burgess reagent (1-methoxy-N-triethylammoniosulfonylmethanimidate, 2.58 g, 10.8 mmol) in toluene (100 mL) was heated at 65° C. for 18 hours, then cooled to room temperature and concentrated in vacuo. The residue was partitioned between ethyl acetate and saturated aqueous sodium bicarbonate solution; the aqueous layer was extracted twice with ethyl acetate, and the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated under reduced pressure. Silica gel chromatography (Gradient: 0% to 100% ethyl acetate in heptane) provided the product as a yellow solid. Yield: 1.0 g, 2.1 mmol, 48%. LCMS m/z 470.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD), observed peaks: δ 8.12 (br d, J=7 Hz, 2H), 7.61 (q, J=1.3 Hz, 1H), 7.42-7.57 (m, 4H), 7.03-7.13 (m, 2H), 4.90 (dd, J=11.8, 2.4 Hz, 1H), 4.30 (dd, J=11.9, 1.6 Hz, 1H), 3.99 (d, J=11.9 Hz, 1H), 2.99 (dd, half of ABX pattern, J=13.2, 4.2 Hz, 1H), 2.81 (dd, half of ABX pattern, J=13.2, 2.8 Hz, 1H), 2.37-2.50 (m, 1H), 2.14 (d, J=1.2 Hz, 3H), 2.01-2.08 (m, 1H).

Step 4. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(4-methyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (4)

A mixture of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(4-methyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C30) (1.0 g, 2.1 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (95%, 0.335 mL, 2.13 mmol) in methanol (34 mL) was heated to 70° C. for 18 hours, then cooled to room temperature and added to an aqueous solution of sodium bicarbonate. The mixture was extracted three times with ethyl acetate, and the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo. Purification was carried out twice using silica gel chromatography (Gradient: 0% to 10% methanol in dichloromethane, followed by a column eluted with ethyl acetate), affording the product as a white solid. Yield: 491 mg, 1.34 mmol, 64%. LCMS m/z 366.1 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 7.60 (q, J=1.3 Hz, 1H), 7.37 (ddd, J=9.6, 8.8, 6.6 Hz, 1H), 6.94-7.03 (m, 2H), 4.80 (dd, J=11.9, 2.5 Hz, 1H), 4.25 (dd, J=11.1, 2.0 Hz, 1H), 3.78 (d, J=11.1 Hz, 1H), 3.00-3.07 (m, 1H), 2.92 (dd, half of ABX pattern, J=12.5, 4.1 Hz, 1H), 2.74 (dd, half of ABX pattern, J=12.6, 2.8 Hz, 1H), 2.32-2.43 (m, 1H), 2.15 (d, J=1.2 Hz, 3H), 1.87 (ddd, J=13.3, 4.0, 2.6 Hz, 1H).

Example 5 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(5-methyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-amine (5)

Step 1. Synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2-oxopropyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C31)

The product was synthesized according to the general procedure for the preparation of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C20) in Example 1, by utilizing a slurry of 1-aminopropan-2-one, hydrochloride salt (340 mg, 3.10 mmol) and N,N-diisopropylethylamine (560 μL, 3.20 mmol) in N,N-dimethylformamide (1 mL) in place of 2,2-dimethoxyethanamine. In this case, the combined organic layers were washed with aqueous hydrochloric acid (1 M, 30 mL) prior to being dried over sodium sulfate. The product was obtained as a very thick oil. Yield: 154 mg, 0.316 mmol, 41%. LCMS m/z 488.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ 8.16-8.28 (br m, 2H), 7.36-7.58 (m, 4H), 4.27-4.37 (m, 1H), 4.22 (apparent br d, J=11.7 Hz, 2H), 3.88-4.04 (m, 2H), 3.12-3.25 (br m, 1H), 3.03 (dd, J=12.9, 3.9 Hz, 1H), 2.19 (s, 3H).

Step 2. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(5-methyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C32)

(4aR,6R,8aS)-2-(Benzoylamino)-8a-(2,4-difluorophenyl)-N-(2-oxopropyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C31) was converted to the product according to the general procedure for the synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(4-methyl-1,3-oxazol-2-yl)-4,4a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C30) in Example 4. In this case, heating was carried out for 2.5 hours, and the work-up consisted of simply concentrating the reaction mixture in vacuo and subjecting it to chromatography on silica gel (Gradient: 0% to 100% ethyl acetate in heptane). The product was obtained as an off-white solid. Yield: 89.9 mg, 0.191 mmol, 64%. ¹H NMR (400 MHz, CD₃OD) δ 8.12 (br d, J=7 Hz, 2H), 7.42-7.57 (m, 4H), 7.03-7.14 (m, 2H), 6.76-6.78 (m, 1H), 4.89 (dd, J=12.0, 2.2 Hz, 1H), 4.30 (br d, J=12.1 Hz, 1H), 3.99 (d, J=11.9 Hz, 1H), 3.25-3.35 (m, 1H, assumed; obscured by solvent peak), 2.99 (dd, J=13.1, 4.1 Hz, 1H), 2.81 (dd, J=13.2, 2.8 Hz, 1H), 2.38-2.50 (m, 1H), 2.31-2.33 (m, 3H), 2.00-2.08 (m, 1H).

Step 3. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(5-methyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (5)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(5-methyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C32) was converted to the product according to the general procedure for the synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (1) in Example 1. The product was obtained as a white solid. Yield: 52.9 mg, 0.145 mmol, 80%. LCMS m/z 366.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 7.37 (ddd, J=9.6, 8.8, 6.6 Hz, 1H), 6.95-7.03 (m, 2H), 6.76-6.78 (m, 1H), 4.79 (dd, J=11.9, 2.3 Hz, 1H), 4.25 (dd, J=11.3, 2.0 Hz, 1H), 3.78 (d, J=11.1 Hz, 1H), 3.00-3.08 (m, 1H), 2.92 (dd, J=12.5, 4.1 Hz, 1H), 2.75 (dd, J=12.6, 2.8 Hz, 1H), 2.33-2.43 (m, 1H), 2.33 (d, J=1.2 Hz, 3H), 1.86 (ddd, J=13.3, 3.9, 2.5 Hz, 1H).

Example 6 (4R,4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-4-methyl-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (6)

Step 1. Synthesis of (4R,4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4-methyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylic acid (C33)

The product, obtained as a pink/white solid, was prepared from N-[(4R,4a R,6R,8aS)-8a-(2,4-difluorophenyl)-6-(hydroxymethyl)-4-methyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (P2) according to the procedure for the synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylic acid (C19) in Example 1. Yield: 2.92 g, 6.54 mmol, 95%. LCMS m/z 447.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 8.08-8.12 (m, 2H), 7.52-7.58 (m, 1H), 7.41-7.49 (m, 3H), 7.02-7.11 (m, 2H), 4.34 (dd, J=12.1, 2.7 Hz, 1H), 4.20 (br d, J=11.9 Hz, 1H), 3.98 (d, J=11.9 Hz, 1H), 3.18-3.26 (m, 1H), 3.06 (ddd, J=12.1, 3.9, 3.9 Hz, 1H), 2.15 (ddd, J=13.6, 4.0, 2.9 Hz, 1H), 1.71-1.83 (m, 1H), 1.27 (d, J=6.8 Hz, 3H).

Step 2. Synthesis of (4R,4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4-methyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C34)

The product, obtained as a white solid, was prepared from (4R,4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4-methyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylic acid (C33) according to the procedure for the synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazine-6-carboxamide (C20) in Example 1. Yield: 418 mg, 0.783 mmol, 81%. LCMS m/z 534.3 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ 8.16-8.28 (m, 2H), 7.34-7.57 (m, 4H), 6.85-6.99 (m, 2H), 6.70-6.78 (m, 1H), 4.37 (t, J=5.4 Hz, 1H), 3.90 (d, J=12.3 Hz, 1H), 3.37 (s, 3H), 3.35 (s, 3H), 2.90-3.00 (m, 1H), 2.27-2.36 (m, 1H), 1.69-1.83 (m, 1H), 1.29 (d, J=7 Hz, 3H).

Step 3. Synthesis of (4R,4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4-methyl-N-(2-oxoethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C35)

The product, obtained as a white solid, was prepared from (4R,4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4-methyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C34) according to the procedure for the synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2-oxoethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C21) in Example 1. Yield: 265 mg, 0.544 mmol, 70%. LCMS m/z 488.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ 9.65 (s, 1H), 8.16-8.26 (m, 2H), 6.85-7.00 (m, 2H), 4.28-4.38 (m, 1H), 4.19-4.27 (m, 2H), 3.99-4.07 (m, 1H), 3.23-3.33 (m, 1H), 2.91-3.02 (m, 1H), 2.27-2.36 (m, 1H), 1.30 (d, J=6 Hz, 3H).

Step 4. Synthesis of N-[(4R,4aR,6R,8aS)-8a-(2,4-difluorophenyl)-4-methyl-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C36)

The product, obtained as a yellow solid, was prepared from (4R,4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4-methyl-N-(2-oxoethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C35) according to the procedure for the synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(4-methyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C30) in Example 4. Yield: 48 mg, 0.10 mmol, 37%. LCMS m/z 470.1 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 8.05-8.14 (m, 2H), 7.92 (d, J=0.8 Hz, 1H), 7.42-7.58 (m, 4H), 7.17 (d, J=0.8 Hz, 1H), 7.02-7.14 (m, 2H), 4.93-4.98 (m, 1H), 4.34 (br d, J=11.9 Hz, 1H), 4.02 (d, J=11.9 Hz, 1H), 3.21-3.29 (br m, 1H), 3.11-3.19 (br m, 1H), 2.10-2.21 (m, 2H), 1.29 (d, J=7.0 Hz, 3H).

Step 5. Synthesis of (4R,4aR,6R,8aS)-8a-(2,4-difluorophenyl)-4-methyl-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (6)

The product, obtained as an off-white solid, was prepared from N-[(4R,4aR,6R,8aS)-8a-(2,4-difluorophenyl)-4-methyl-6-(1,3-oxazol-2-yl)-4,4a,5,6,8,8a-hexahydro pyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C36) according to the procedure for the synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (1) in Example 1. Yield: 15.6 mg, 42.7 μmol, 84%. LCMS m/z 366.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 7.91-7.92 (m, 1H), 7.31-7.40 (m, 1H), 7.16-7.18 (m, 1H), 6.94-7.04 (m, 2H), 4.83-4.89 (m, 1H, assumed; partially obscured by water peak), 4.29 (br d, J=11.3 Hz, 1H), 3.84 (d, J=11.1 Hz, 1H), 3.12-3.20 (m, 1H), 2.91 (ddd, J=11.9, 3.7, 3.5 Hz, 1H), 2.01-2.12 (m, 1H), 1.93-2.00 (m, 1H), 1.21 (d, J=6.9 Hz, 3H).

Example 7A (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (7)

Step 1. Synthesis of [(4aR,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4-a,5,8,8a-tetrahydropyrano[3,4-][1,3]thiazin-6(4H)-ylidene]methyl acetate (C37)

Acetic anhydride (1.5 mL, 16 mmol) was added to a slurry of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-formyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-c][1,3]thiazin-2-yl]benzamide (C23) (661 mg, 1.59 mmol) and potassium carbonate (1.34 g, 9.70 mmol) in acetonitrile (16 mL). After the flask had been flushed with nitrogen, the reaction mixture was heated at reflux for 2.5 hours, then allowed to cool to room temperature and stir for 18 hours. The slurry was diluted with ethyl acetate and filtered; the solids were washed with ethyl acetate, and the combined filtrates were concentrated in vacuo. Purification using silica gel chromatography (Gradient: 0% to 100% ethyl acetate in heptane) afforded the product as a white solid, which was assigned as a roughly 4:1 mixture of geometric isomers from the ¹H NMR. Yield: 437 mg, 0.953 mmol, 60%. LCMS m/z 459.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), major isomer only: δ 8.09-8.32 (br s, 2H), 7.50-7.56 (m, 1H), 7.39-7.45 (m, 3H), 6.85-6.99 (m, 2H), 6.75 (d, J=1.9 Hz, 1H), 4.31 (dd, J=11.7, 1.2 Hz, 1H), 4.02 (d, J=11.8 Hz, 1H), 3.13-3.26 (br s, 1H), 2.97-3.07 (m, 1H), 2.70-2.87 (m, 2H), 2.19 (s, 3H), 2.17-2.25 (m, 1H).

Step 2. Synthesis of N-[(4aR,8aS)-8a-(2,4-difluorophenyl)-6-oxo-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C38)

A solution of [(4aR,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4-a,5,8,8a-tetrahydropyrano[3,4-][1,3]thiazin-6(4H)-ylidene]methyl acetate (C37) (430 mg, 0.938 mmol), ruthenium(III) chloride (5.8 mg, 28 μmol) and sodium periodate (98.5%, 407 mg, 1.87 mmol) in acetonitrile (0.5 mL) and a 1:1 mixture of 1,2-dichloroethane and water (5 mL) was stirred for 3 hours at room temperature, then allowed to stand for 18 hours without stirring. After dilution with saturated aqueous sodium thiosulfate solution (25 mL), the mixture was extracted with ethyl acetate (3×50 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (Gradient: 0% to 80% ethyl acetate in heptane) provided the product as a white solid. Yield: 237 mg, 0.589 mmol, 63%. LCMS m/z 403.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.99 (br d, J=8 Hz, 2H), 7.49-7.54 (m, 1H), 7.43 (br dd, J=8, 7 Hz, 2H), 7.32 (ddd, J=9.0, 9.0, 6.3 Hz, 1H), 6.81-6.93 (m, 2H), 4.85 (d, J=11.7 Hz, 1H), 4.24 (d, J=11.5 Hz, 1H), 3.35-3.44 (m, 1H), 2.87-2.97 (m, 2H), 2.80 (dd, half of ABX pattern, J=18.7, 7.5 Hz, 1H), 2.63 (dd, J=13.1, 3.1 Hz, 1H).

Step 3. Synthesis of N-[(4aR,8aS)-8a-(2,4-difluorophenyl)-6-hydroxy-6-(1-methyl-1H-pyrazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C39)

n-Butyllithium (2.5 M solution in hexanes, 0.518 mL, 1.30 mmol) was added drop-wise to a 78° C. solution of 4-bromo-1-methyl-1H-pyrazole (98%, 0.155 mL, 1.47 mmol) in tetrahydrofuran (4 mL). The resulting slurry was stirred at 78° C. for 30 minutes. A solution of N-[(4aR,8aS)-8a-(2,4-difluorophenyl)-6-oxo-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C38) (237 mg, 0.589 mmol) in tetrahydrofuran (1.5 mL) was then added drop-wise, and the reaction mixture was allowed to stir for 2.5 hours at 78° C. The reaction was quenched with saturated aqueous ammonium chloride solution (15 mL) and allowed to warm slowly to room temperature, at which point it was extracted with ethyl acetate (3×25 mL), dried over sodium sulfate, filtered, and concentrated in vacuo to provide the crude product as a foam (317 mg), which was taken directly to the following step. LCMS m/z 485.2 [M+H⁺].

Step 4. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-4,4a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C40)

Triethylsilane (99%, 1.42 mL, 8.81 mmol) was added to a solution of N-[(4aR,8aS)-8a-(2,4-difluorophenyl)-6-hydroxy-6-(1-methyl-1H-pyrazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C39) (317 mg from the previous step, ≦0.589 mmol) in dichloromethane (4 mL), and the resulting mixture was cooled to 15° C. Trimethylsilyl trifluoromethanesulfonate (0.429 mL, 2.35 mmol) was added drop-wise to the reaction mixture over 5 to 10 minutes, at which time the solution was allowed to warm to room temperature and stir for 30 minutes. The flask was cooled in an ice bath, and triethylsilane (99%, 1.0 mL, 6.2 mmol) and trifluoroacetic acid (1 mL) were added; the reaction mixture was then allowed to warm to room temperature. After 45 minutes, the reaction mixture was concentrated in vacuo, then carefully treated with saturated aqueous sodium bicarbonate solution to neutralize remaining acid. After extraction with ethyl acetate (3×25 mL), the combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was slurried in heptane and filtered; the isolated solid, a mixture of diastereomers, was purified using silica gel chromatography (10% to 100% ethyl acetate in heptane), providing the product (lower R_(f) material, major isomer, 77 mg) and mixed fractions (33 mg). The mixed material was subjected to preparative thin layer chromatography (Eluent: 3:2 ethyl acetate/heptane) to afford additional product. Total yield: 107 mg, 0.228 mmol, 39%. LCMS m/z 469.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.18-8.24 (m, 2H), 7.48-7.53 (m, 1H), 7.40-7.46 (m, 4H), 7.39 (s, 1H), 6.85-6.96 (m, 2H), 4.71 (dd, J=11.5, 2.2 Hz, 1H), 4.28 (dd, J=12.2, 1.8 Hz, 1H), 3.84 (s, 3H), 3.83-3.89 (m, 1H), 3.20-3.29 (m, 1H), 3.03 (dd, J=12.9, 4.1 Hz, 1H), 2.67 (dd, J=12.9, 2.8 Hz, 1H), 2.22-2.33 (m, 1H), 1.92 (ddd, J=13.7, 4.0, 2.4 Hz, 1H). The relative and absolute stereochemistry of this material was confirmed via X-ray crystallography:

Single Crystal X-Ray Analysis

Data collection was performed on a Bruker APEX diffractometer at room temperature. Data collection consisted of 3 omega scans at low angle and three at high angle, each with 0.5 step. In addition, 2 high angle phi scans were collected to improve the quality of the absorption correction.

The structure was solved by direct methods using the SHELX software suite in space group P2(1). The structure was subsequently refined by the full-matrix least squares method. All non-hydrogen atoms were found and refined using anisotropic displacement parameters.

The hydrogen atoms located on nitrogen were found from the Fourier difference map and refined with distance restrained. The remaining hydrogen atoms were placed in calculated positions and were allowed to ride on their carrier atoms. The final refinement included isotropic displacement parameters for all hydrogen atoms.

Analysis of the absolute structure using likelihood methods (Hooft, 2008) was performed using PLATON (Spek, 2010). The method calculates that the probability that the structure is correct is 100.0%. The Hooft parameter is reported as 0.05 with an esd of 0.018.

The final R-index was 5.9%. A final difference Fourier revealed no missing or misplaced electron density.

Pertinent crystal, data collection and refinement are summarized in Table 1. Atomic coordinates, bond lengths, bond angles, torsion angles and displacement parameters are listed in Tables 2-5.

SOFTWARE AND REFERENCES

-   SHELXTL, Version 5.1, Bruker AXS, 1997. -   PLATON, A. L. Spek, J. Appl. Cryst. 2003, 36, 7-13. -   MERCURY, C. F. Macrae et al., J. Appl. Cryst. 2006, 39, 453-457. -   R. W. W. Hooft et al., J. Appl. Cryst. 2008, 41, 96-103. -   H. D. Flack, Acta Cryst. 1983, A39, 867-881.

TABLE 1 Crystal data and structure refinement for N-[(4aR,6R,8aS)—8a- (2,4-difluorophenyl)—6-(1-methyl-1H-pyrazol-4-yl)—4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C40). Empirical formula C24 H22 F2 N4 O2 S Formula weight 468.52 Temperature 296(2)K Wavelength 1.54178 Å Crystal system Monoclinic Space group P2(1) Unit cell dimensions a = 9.8090(2) Å α = 90°. b = 12.3169(2) Å β = 117.9320(10)°. c = 10.1983(2) Å γ = 90°. Volume 1088.59(4) Å³ Z 2 Density (calculated) 1.429 Mg/m³ Absorption coefficient 1.734 mm⁻¹ F(000) 488 Crystal size 0.21 × 0.16 × 0.14 mm³ Theta range for data collection 4.91 to 69.41°. Index ranges −11 ≦ h ≦ 10, −13 ≦ k ≦ 14, −12 ≦ l ≦ 12 Reflections collected 5220 Independent reflections 3272 [R(int) = 0.0468] Completeness to theta = 69.41° 93.7% Absorption correction Empirical Refinement method Full-matrix least-squares on F² Data/restraints/parameters 3272/2/302 Goodness-of-fit on F² 1.051 Final R indices [l > 2sigma(l)] R1 = 0.0592, wR2 = 0.1458 R indices (all data) R1 = 0.0610, wR2 = 0.1482 Absolute structure parameter 0.02(2) Largest diff. peak and hole 0.399 and −0.584 e.Å⁻³

TABLE 2 Atomic coordinates (×10⁴) and equivalent isotropic displacement parameters (Å² × 10³ ) for C40. U(eq) is defined as one third of the trace of the orthogonalized U^(ij) tensor. x y z U(eq) C(1) 2513(5) 8761(4) 7563(5) 58(1) C(2) 2222(7) 9560(5) 8326(6) 76(1) C(3) 1209(6) 10397(5)  7575(8) 82(2) C(4)  493(6) 10422(5)  6037(7) 82(2) C(5)  776(5) 9612(4) 5289(6) 65(1) C(6) 1793(4) 8771(3) 6026(4) 48(1) C(7) 2218(4) 7939(3) 5231(4) 45(1) C(8) 2022(4) 7562(3) 2909(3) 39(1) C(9) 4177(3) 6703(3) 2603(3) 36(1) C(10) 2997(4) 6202(3) 1117(3) 41(1) C(11) 1821(4) 7048(3)  153(3) 52(1) C(12) 5094(3) 7650(3) 2432(3) 40(1) C(13) 5821(4) 7601(3) 1545(3) 46(1) C(14) 6699(5) 8439(4) 1423(4) 58(1) C(15) 6840(5) 9370(4) 2226(4) 56(1) C(16) 6167(5) 9470(4) 3123(4) 58(1) C(17) 5315(4) 8606(3) 3233(3) 47(1) C(18) 2295(4) 5171(3) 1385(3) 42(1) C(19) 3565(4) 4376(3) 2361(3) 42(1) C(20) 5321(3) 5806(3) 3542(3) 39(1) C(21) 2926(4) 3343(3) 2621(3) 42(1) C(22) 3185(5) 2315(3) 2246(4) 49(1) C(23) 2113(6)  468(4) 2236(7) 82(2) C(24) 1865(4) 3191(3) 3131(4) 51(1) F(1) 5691(3) 6699(2)  728(3) 70(1) F(2) 7668(3) 10193(2)  2093(3) 81(1) N(1) 1442(3) 8014(2) 3712(3) 46(1) N(2) 3367(3) 7038(2) 3435(3) 38(1) N(3) 2318(4) 1625(3) 2545(3) 52(1) N(4) 1492(4) 2132(3) 3105(4) 57(1) O(1) 3201(3) 7245(2) 5942(3) 58(1) O(2) 4569(3) 4887(2) 3759(2) 42(1) S(1)  800(1) 7697(1) 1003(1) 54(1)

TABLE 3 Bond lengths [Å] and angles [°] for C40. C(1)—C(2) 1.365(6) C(2)—C(1)—C(6) 120.5(4) C(1)—C(6) 1.385(6) C(1)—C(2)—C(3) 120.5(5) C(2)—C(3) 1.387(8) C(4)—C(3)—C(2) 119.6(5) C(3)—C(4) 1.386(9) C(5)—C(4)—C(3) 119.3(5) C(4)—C(5) 1.362(7) C(4)—C(5)—C(6) 121.7(5) C(5)—C(6) 1.389(6) C(1)—C(6)—C(5) 118.4(4) C(6)—C(7) 1.483(5) C(1)—C(6)—C(7) 119.2(3) C(7)—O(1) 1.237(4) C(5)—C(6)—C(7) 122.2(4) C(7)—N(1) 1.372(4) O(1)—C(7)—N(1) 125.2(3) C(8)—N(1) 1.319(4) O(1)—C(7)—C(6) 119.9(3) C(8)—N(2) 1.335(4) N(1)—C(7)—C(6) 114.9(3) C(8)—S(1) 1.749(3) N(1)—C(8)—N(2) 126.0(3) C(9)—N(2) 1.467(4) N(1)—C(8)—S(1) 112.3(2) C(9)—C(12) 1.532(5) N(2)—C(8)—S(1) 121.7(2) C(9)—C(10) 1.541(4) N(2)—C(9)—C(12) 110.9(3) C(9)—C(20) 1.546(4) N(2)—C(9)—C(10) 108.7(3) C(10)—C(11) 1.523(5) C(12)—C(9)—C(10) 113.8(2) C(10)—C(18) 1.529(5) N(2)—C(9)—C(20) 106.0(2) C(11)—S(1) 1.791(4) C(12)—C(9)—C(20) 108.7(2) C(12)—C(17) 1.390(5) C(10)—C(9)—C(20) 108.4(3) C(12)—C(13) 1.390(5) C(11)—C(10)—C(18) 114.5(3) C(13)—F(1) 1.358(5) C(11)—C(10)—C(9) 110.7(3) C(13)—C(14) 1.386(5) C(18)—C(10)—C(9) 110.5(2) C(14)—C(15) 1.378(6) C(10)—C(11)—S(1) 114.8(2) C(15)—F(2) 1.345(5) C(17)—C(12)—C(13) 115.6(3) C(15)—C(16) 1.361(6) C(17)—C(12)—C(9) 121.1(3) C(16)—C(17) 1.390(5) C(13)—C(12)—C(9) 123.3(3) C(18)—C(19) 1.530(4) F(1)—C(13)—C(14) 115.9(3) C(19)—O(2) 1.444(4) F(1)—C(13)—C(12) 120.3(3) C(19)—C(21) 1.495(5) C(14)—C(13)—C(12) 123.7(4) C(20)—O(2) 1.424(4) C(15)—C(14)—C(13) 117.4(3) C(21)—C(24) 1.378(5) F(2)—C(15)—C(16) 120.4(4) C(21)—C(22) 1.381(5) F(2)—C(15)—C(14) 117.7(4) C(22)—N(3) 1.335(5) C(16)—C(15)—C(14) 121.9(4) C(23)—N(3) 1.452(6) C(15)—C(16)—C(17) 119.0(4) C(24)—N(4) 1.350(5) C(16)—C(17)—C(12) 122.4(3) N(3)—N(4) 1.344(5) C(10)—C(18)—C(19) 110.5(3) O(2)—C(19)—C(18) 109.5(3) C(8)—N(1)—C(7) 120.0(3) C(21)—C(19)—C(18) 112.3(3) C(8)—N(2)—C(9) 127.2(3) O(2)—C(20)—C(9) 112.7(2) C(22)—N(3)—N(4) 112.1(3) C(24)—C(21)—C(22) 104.4(3) C(22)—N(3)—C(23) 127.8(4) 24)—C(21)—C(19) 129.5(3) N(4)—N(3)—C(23) 119.9(4) C(22)—C(21)—C(19) 125.8(3) N(3)—N(4)—C(24) 104.2(3) N(3)—C(22)—C(21) 107.5(3) C(20)—O(2)—C(19) 110.9(2) N(4)—C(24)—C(21) 111.7(4) C(8)—S(1)—C(11) 104.37(15)

Symmetry Transformations Used to Generate Equivalent Atoms

TABLE 4 Anisotropic displacement parameters (Å² × 10³) for C40. The anisotropic displacement factor exponent takes the form: −2π²[h² a*²U¹¹ + . . . + 2 h k a* b* U¹²] U¹¹ U²² U³³ U²³ U¹³ U¹² C(1)  75(3) 46(2) 74(2)    3(2) 52(2)  −3(2) C(2)  104(4)  61(3) 92(3) −14(3) 71(3) −16(3) C(3)  88(3) 54(3) 138(5)  −35(3) 81(3) −17(2) C(4)  63(3) 57(3) 126(4)  −13(3) 45(3)   15(2) C(5)  52(2) 54(3) 89(3)  −9(2) 32(2)    8(2) C(6)  48(2) 37(2) 73(2)  −3(2) 39(2)  −4(1) C(7)  42(2) 36(2) 61(2)    2(1) 27(1)  −1(1) C(8)  35(2) 27(2) 50(1)    2(1) 17(1)  −2(1) C(9)  38(2) 33(2) 35(1)    0(1) 17(1)    0(1) C(10) 45(2) 38(2) 35(1)    0(1) 15(1)  −3(1) C(11) 59(2) 43(2) 38(1)    8(1)  9(1)  −5(2) C(12) 39(2) 40(2) 37(1)    3(1) 14(1)  −1(1) C(13) 56(2) 41(2) 47(1)  −1(2) 28(1)  −2(2) C(14) 61(2) 66(3) 52(2)    9(2) 30(2) −10(2) C(15) 56(2) 53(2) 51(2)    9(2) 17(2) −15(2) C(16) 67(2) 41(2) 60(2)  −8(2) 24(2) −14(2) C(17) 51(2) 43(2) 44(2)  −5(1) 21(1)  −8(2) C(18) 46(2) 33(2) 39(1)  −4(1) 13(1)  −5(1) C(19) 47(2) 34(2) 45(1)  −4(1) 20(1)  −3(1) C(20) 40(2) 34(2) 40(1)  −2(1) 14(1)  1(1) C(21) 47(2) 30(2) 45(1)    0(1) 18(1)  −2(1) C(22) 53(2) 37(2) 53(2)    0(1) 21(2)    5(1) C(23) 84(3) 36(2) 107(4)   −5(2) 28(3)  −3(2) C(24) 50(2) 41(2) 59(2)  −3(2) 24(2)  −2(2) F(1)  92(2) 56(2) 88(2) −18(1) 64(1) −14(1) F(2)  90(2) 71(2) 75(1)   13(1) 32(1) −37(1) N(1)  42(2) 37(2) 59(2)    3(1) 23(1)    5(1) N(2)  40(1) 36(2) 38(1)    1(1) 17(1)    4(1) N(3)  58(2) 31(2) 54(2)    3(1) 14(1)    0(1) N(4)  54(2) 45(2) 67(2)    4(2) 24(2)  −2(1) O(1)  73(2) 49(2) 56(1)    9(1) 33(1)   21(1) O(2)  47(1) 35(1) 39(1)    1(1) 16(1)  −3(1) S(1)  48(1) 45(1) 50(1)    9(1)  7(1)    8(1)

TABLE 5 Hydrogen coordinates (×10⁴) and isotropic displacement parameters (Å² × 10³ ) for C40. x y z U(eq) H(1) 3199 8206 8078 70 H(2) 2706 9542 9358 91 H(3) 1012 10938 8099 99 H(4) −174 10986 5522 98 H(5) 276 9622 4257 78 H(10) 3570 5982 590 49 H(11A) 1078 6700 −756 63 H(11B) 2351 7600 −116 63 H(14) 7174 8375 823 70 H(16) 6274 10107 3653 70 H(17) 4878 8668 3865 56 H(18A) 1625 4830 442 51 H(18B) 1676 5358 1867 51 H(19) 4176 4198 1857 51 H(20A) 5913 5575 3051 47 H(20B) 6036 6104 4501 47 H(22) 3844 2135 1856 59 H(23A) 1552 355 1183 123 H(23B) 1547 161 2701 123 H(23C) 3104 124 2621 123 H(24) 1455 3748 3454 61 H(99A) 3780(40) 6790(30) 4420(20) 46(10)

Step 5. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (7)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C40) was converted to the product according to the general procedure for the synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (1) in Example 1. The product was obtained as a colorless foam. Yield: 14.5 mg, 39.8 μmol, 92%. LCMS m/z 365.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.48 (s, 1H), 7.42 (s, 1H), 7.37 (ddd, J=9, 9, 6.8 Hz, 1H), 6.78-6.92 (m, 2H), 4.68 (dd, J=11.3, 2.0 Hz, 1H), 4.22 (dd, J=11.2, 1.9 Hz, 1H), 3.88 (s, 3H), 3.86-3.91 (m, 1H), 2.97-3.07 (m, 2H), 2.61-2.68 (m, 1H), 2.07-2.19 (m, 1H), 1.74-1.82 (m, 1H).

Example 7B

Alternate synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (7)

Step 1. Synthesis of N-{(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(2-methoxyethenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl}benzamide (C41)

Commercial methoxymethyl(triphenyl)phosphonium chloride (9.49 g) was placed in a vacuum oven at 80° C. and low vacuum for 18 hours. The resulting material (9.38 g, 27.4 mmol) was suspended in tetrahydrofuran (130 mL) and cooled to an internal temperature of 2° C. A solution of potassium tert-butoxide (1.0 M in tetrahydrofuran, 23.5 mL, 23.5 mmol) was added over 2 to 3 minutes, at a rate that kept the reaction temperature under 5° C. The resulting solution was stirred for 5 minutes at 2 to 5° C., then warmed to room temperature over 20 minutes. The reaction mixture was cooled to 3° C. internal temperature and treated with a solution of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-formyl-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C23) (3.20 g, 7.68 mmol) in tetrahydrofuran (20 mL) over 2 to 3 minutes, while keeping the internal temperature below 6° C. After 15 minutes at 3 to 6° C., the reaction mixture was allowed to warm to room temperature over 25 minutes, then cooled to 14° C. and quenched by addition of saturated aqueous sodium bicarbonate solution (125 mL). The resulting mixture was extracted with ethyl acetate (3×150 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. Purification via silica gel chromatography (Gradient: 0% to 35% ethyl acetate in heptane) provided the product as a white solid, which was characterized by ¹H NMR as a roughly 1.4:1 mixture of geometrical isomers around the double bond. Yield: 2.66 g, 5.98 mmol, 78%. LCMS m/z 445.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.24 (br d, J=7.3 Hz, 2H), 7.48-7.54 (m, 1H), 7.37-7.48 (m, 3H), 6.85-6.96 (m, 2H), [6.68 (d, J=12.7 Hz) and 5.99 (dd, J=6.3, 0.9 Hz), total 1H], [4.86 (dd, J=12.7, 8.0 Hz) and 4.55 (dd, half of ABX pattern, J=8.1, 6.3 Hz), total 1H], 4.60-4.68 and 4.06-4.13 (2 m, total 1H), [4.20 (dd, J=12.2, 1.9 Hz) and 4.19 (dd, J=12.2, 2.0 Hz), total 1H], [3.81 (d, J=12.2 Hz) and 3.79 (d, J=12.2 Hz), total 1H], 3.66 and 3.55 (2 s, total 3H), 3.13-3.24 (m, 1H), 2.97-3.05 (m, 1H), 2.60-2.68 (m, 1H), 1.97-2.19 (m, 1H), 1.66-1.75 (m, 1H).

Step 2. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,1,3,3-tetramethoxypropan-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C42)

To a stirring mixture of N-{(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(2-methoxyethenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl}benzamide (C41) (70.0 mg, 0.157 mmol) in 1,2-dichloroethane (150 μL) was added trimethyl orthoformate (36.1 μL, 0.330 mmol), followed by boron trifluoride diethyl etherate (7.0 μL, 56 μmol), and the reaction mixture was stirred at room temperature for 30 minutes. At this point, additional trimethyl orthoformate (17 μL, 0.15 mmol), and boron trifluoride diethyl etherate (7.0 μL, 56 μmol) were introduced, and stirring was continued for 30 minutes. Boron trifluoride diethyl etherate (7.0 μL, 56 μmol) was added again, and the reaction mixture was stirred for 1 hour, then diluted with dichloromethane (1 mL) and transferred via pipette into a stirring mixture of saturated aqueous sodium bicarbonate solution (2 mL) and dichloromethane (5 mL). The aqueous layer was extracted with dichloromethane (5 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to afford the product as an off-white to pale yellow foam. Yield: 75 mg, 0.14 mmol, 89%. LCMS m/z 549.1 [M−H⁺]. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ 8.22-8.26 (m, 2H), 7.37-7.53 (m, 4H), 6.84-6.95 (m, 2H), 4.56 (d, J=4.9 Hz, 1H), 4.52 (d, J=3.8 Hz, 1H), 4.14 (dd, J=12.2, 1.8 Hz, 1H), 4.03-4.09 (m, 1H), 3.78 (d, J=12.2 Hz, 1H), 3.44 (s, 3H), 3.41 (s, 3H), 3.40 (s, 3H), 3.36 (s, 3H), 2.98 (dd, J=12.8, 4.2 Hz, 1H), 2.65 (dd, J=12.8, 2.7 Hz, 1H), 2.22-2.33 (m, 1H), 2.13-2.17 (m, 1H), 1.77 (ddd, J=14, 4, 2 Hz, 1H).

Step 3. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (7)

Water (100 μL) was added to a mixture of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,1,3,3-tetramethoxypropan-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C42) (70 mg, 0.13 mmol) in methanol (150 μL). To the resulting gel was added methylhydrazine (10 μL, 0.19 mmol) followed by concentrated sulfuric acid (13 μL, 0.24 mmol). The mixture was vortexed for 2 to 3 minutes to break up the gel, then heated to 45° C. for 2.5 hours and to 60° C. for 20 hours. After cooling to room temperature, the reaction mixture was partitioned between water (5 mL) and dichloromethane (2 mL). The aqueous layer was adjusted to pH 8-9 via drop-wise addition of 1 M aqueous sodium hydroxide solution, then extracted with ethyl acetate (2×10 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. Purification via silica gel chromatography (Gradient: 0% to 18% methanol in dichloromethane) provided the product as an off-white solid. Yield: 19 mg, 52 μmol, 40%. LCMS m/z 365.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.48 (s, 1H), 7.42 (s, 1H), 7.38 (ddd, J=9.0, 9.0, 6.6 Hz, 1H), 6.85-6.91 (m, 1H), 6.82 (ddd, J=12.5, 8.6, 2.5 Hz, 1H), 4.67 (dd, J=11.5, 2.2 Hz, 1H), 4.22 (dd, J=11.1, 2.4 Hz, 1H), 3.88 (s, 3H), 3.84-3.89 (m, 1H), 2.96-3.05 (m, 2H), 2.60-2.67 (m, 1H), 2.07-2.19 (m, 1H), 1.73-1.80 (m, 1H).

Example 8 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(4-methyl-1,3-thiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (8)

Step 1. Synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C43)

The product, obtained as a white solid, was synthesized according to the general procedure for preparation of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C20) in Example 1, except that a 0.5 M solution of ammonia in 1,4-dioxane was used in place of 2,2-dimethoxyethanamine. Yield: 638 mg, 1.48 mmol, 83%. LCMS m/z 432.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.16-8.33 (br m, 2H), 7.35-7.60 (m, 4H), 6.86-7.00 (m, 2H), 6.48-6.59 (br s, 1H), 5.37-5.46 (br s, 1H), 4.16-4.26 (m, 2H), 3.90 (br d, J=12 Hz, 1H), 3.14-3.25 (br m, 1H), 3.04 (dd, J=12.9, 3.9 Hz, 1H), 2.71 (br d, J=13 Hz, 1H), 2.19-2.30 (m, 1H), 2.06-2.19 (br m, 1H).

Step 2. Synthesis of (4aR,6R,8aS)-2-amino-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C44)

A mixture of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C43) (630 mg, 1.46 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (95%, 0.230 mL, 1.46 mmol) in methanol (18 mL) was heated to 68° C. for 2 hours, treated with additional 1,8-diazabicyclo[5.4.0]undec-7-ene (95%, 0.20 mL, 1.3 mmol), then heated for 18 hours. The reaction mixture was allowed to cool to room temperature and concentrated in vacuo. Purification by silica gel chromatography [Gradient: 0% to 100% (89:10:1 dichloromethane/methanol/concentrated ammonium hydroxide) in dichloromethane] afforded the product as a white solid. Yield: 391 mg, 1.19 mmol, 82%. ¹H NMR (400 MHz, CD₃OD) δ 7.33-7.41 (m, 1H), 6.94-7.02 (m, 2H), 4.14 (br d, J=11.2 Hz, 1H), 4.09 (dd, J=11.9, 2.9 Hz, 1H), 3.77 (d, J=11.3 Hz, 1H), 2.92-3.00 (m, 1H), 2.89 (dd, J=12.5, 4.1 Hz, 1H), 2.72 (dd, J=12.5, 2.5 Hz, 1H), 1.91-1.98 (m, 1H), 1.78-1.89 (m, 1H).

Step 3. Synthesis of tert-butyl [(4aR,6R,8aS)-6-carbamoyl-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]carbamate (C45)

A solution of (4aR,6R,8aS)-2-amino-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazine-6-carboxamide (C44) (326 mg, 0.996 mmol) and di-tert-butyl dicarbonate (283 mg, 1.30 mmol) in tetrahydrofuran (30 mL) was allowed to stir at room temperature for 18 hours. After removal of solvent in vacuo, the residue was partitioned between water (15 mL) and ethyl acetate (15 mL). The aqueous layer was extracted with ethyl acetate (2×15 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated under reduced pressure. Chromatography on silica gel (Gradient: 20% to 100% ethyl acetate in heptane) provided the product as a white solid. Yield: 424 mg, 0.992 mmol, 99.6%. LCMS m/z 426.2 [M−H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 7.36-7.43 (m, 1H), 6.99-7.09 (br m, 2H), 4.10-4.18 (br m, 2H), 3.91 (d, J=11.7 Hz, 1H), 3.05-3.18 (br m, 1H), 2.91 (dd, J=13.0, 3.7 Hz, 1H), 2.74 (br d, J=13 Hz, 1H), 2.04 (br d, J=13 Hz, 1H), 1.78-1.90 (m, 1H), 1.51 (s, 9H).

Step 4. Synthesis of tert-butyl [(4aR,6R,8aS)-6-carbamothioyl-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]carbamate (C46)

A mixture of tert-butyl [(4aR,6R,8aS)-6-carbamoyl-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]carbamate (C45) (420 mg, 0.983 mmol) and 2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane-2,4-dithione (Lawesson's reagent, 398 mg, 0.983 mmol) in tetrahydrofuran (20 mL) was stirred at 40° C. for 45 minutes, then allowed to cool to room temperature. After removal of solvent in vacuo, purification was carried out by silica gel chromatography (Gradient: 0% to 100% ethyl acetate in heptane). The product was obtained as a white solid. Yield: 318 mg, 0.717 mmol, 73%. ¹H NMR (400 MHz, CD₃OD) δ 7.40 (ddd, J=9.6, 8.7, 6.5 Hz, 1H), 6.99-7.09 (m, 2H), 4.42 (br d, J=11.5 Hz, 1H), 4.18 (d, J=11.7 Hz, 1H), 3.94 (d, J=11.5 Hz, 1H), 3.10-3.22 (br m, 1H), 2.91 (dd, J=12.9, 3.9 Hz, 1H), 2.74 (dd, J=13.1, 2.3 Hz, 1H), 2.41 (br d, J=12.7 Hz, 1H), 1.70-1.83 (m, 1H), 1.51 (s, 9H).

Step 5. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(4-methyl-1,3-thiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (8)

tert-Butyl [(4aR,6R,8aS)-6-carbamothioyl-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]carbamate (C46) (93 mg, 0.21 mmol) and chloroacetone (84 μL, 1.05 mmol) were combined in toluene (4 mL) and heated at 65° C. for 45 minutes, then at 85° C. for 3 hours. After the reaction mixture had been allowed to cool to room temperature, it was partitioned between saturated aqueous sodium bicarbonate solution (10 mL) and ethyl acetate (15 mL). The aqueous layer was extracted with ethyl acetate (2×15 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. Chromatography on silica gel (Gradient: 0% to 100% ethyl acetate in heptane) afforded the product as a white solid. Yield: 15.6 mg, 40.9 μmol, 19%. LCMS m/z 382.1 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 7.38 (ddd, J=9.6, 8.8, 6.6 Hz, 1H), 7.07-7.09 (m, 1H), 6.95-7.04 (m, 2H), 4.95 (dd, J=10.8, 3.4 Hz, 1H), 4.30 (dd, J=11.2, 2.0 Hz, 1H), 3.85 (d, J=11.3 Hz, 1H), 3.06-3.14 (m, 1H), 2.92 (dd, J=12.5, 4.1 Hz, 1H), 2.75 (dd, J=12.6, 2.8 Hz, 1H), 2.41 (d, J=1.0 Hz, 3H), 1.97-2.13 (m, 2H).

Example 9 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(2-methylpyrimidin-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (9)

Step 1. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(2-methylpyrimidin-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C47)

Water (5 μL, 0.28 mmol) was added to a stirring mixture of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(prop-2-ynoyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C25) (44 mg, 0.10 mmol), acetamidine hydrochloride (12.0 mg, 0.127 mmol) and sodium carbonate (29.0 mg, 0.274 mmol) in ethyl acetate (1 mL) in a vial. The vial was tightly capped and heated to 80° C. for 30 minutes. After cooling to room temperature, the reaction mixture was partitioned between water (2 mL) and ethyl acetate (5 mL). The aqueous layer was extracted with ethyl acetate (6 mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. Chromatography on silica gel (Gradient: 40% to 100% ethyl acetate in heptane) provided the product as an off-white to pale tan foam. Yield: 36 mg, 75 μmol, 75%. LCMS m/z 481.2 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 8.64 (d, J=5.3 Hz, 1H), 8.12 (br d, J=7.2 Hz, 2H), 7.41-7.57 (m, 5H), 7.04-7.15 (m, 2H), 4.78 (br dd, J=11.4, 2 Hz, 1H), 4.34 (d, J=12.3 Hz, 1H), 4.10 (d, J=12.1 Hz, 1H), 3.33-3.42 (br m, 1H), 3.01 (dd, half of ABX pattern, J=13.1, 4.1 Hz, 1H), 2.82 (dd, half of ABX pattern, J=13.2, 2.6 Hz, 1H), 2.67 (s, 3H), 2.23-2.31 (m, 1H), 1.89-2.01 (m, 1H).

Step 2. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(2-methylpyrimidin-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (9)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(2-methylpyrimidin-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C47) was converted to the product according to the method described for synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-3-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (2) in Example 2. The product was obtained as a white solid; the relative stereochemistry of the pyrimidine side chain was confirmed via nuclear Overhauser enhancement study. Yield: 17.9 mg, 47.6 μmol, 71%. LCMS m/z 377.1 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 8.65 (d, J=5.3 Hz, 1H), 7.48 (br d, J=5.3 Hz, 1H), 7.39 (ddd, J=9.5, 8.8, 6.6 Hz, 1H), 6.96-7.04 (m, 2H), 4.69 (dd, J=11.6, 2.6 Hz, 1H), 4.28 (dd, J=11.2, 2.0 Hz, 1H), 3.89 (d, J=11.2 Hz, 1H), 3.08-3.15 (m, 1H), 2.93 (dd, J=12.6, 4.2 Hz, 1H), 2.74 (dd, J=12.6, 2.8 Hz, 1H), 2.68 (s, 3H), 2.09 (ddd, J=13.2, 3.9, 2.7 Hz, 1H), 1.78-1.89 (m, 1H).

Example 10 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-[1-(propan-2-yl)-1H-pyrazol-3-yl]-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (10)

Step 1. Synthesis of N-{(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-[1-(propan-2-yl)-1H-pyrazol-3-yl]-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl}benzamide (C48) and N-{(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-[1-(propan-2-yl)-1H-pyrazol-5-yl]-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl}benzamide (C49)

N,N-Diisopropylethylamine (0.02 mL, 0.11 mmol) was added to a slurry of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(prop-2-ynoyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C25) (34.5 mg, 78.0 μmol) and isopropylhydrazine hydrochloride (8.6 mg, 78 μmol) in 2-propanol (2 mL), and the reaction mixture was stirred at room temperature for 3 hours. After concentration in vacuo, purification was effected via chromatography on silica gel (Gradient: 0% to 60% ethyl acetate in heptane) to provide the product as a gum. This material existed as a roughly 4:1 mixture of isomers via ¹H NMR analysis. Yield: 36 mg, 72 μmol, 92%. LCMS m/z 497.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), major isomer (C48) peaks only: δ 8.23 (br d, J=7 Hz, 2H), 7.40-7.56 (m, 4H), 7.36 (d, J=2.3 Hz, 1H), 6.86-6.99 (m, 2H), 6.30 (br d, J=2 Hz, 1H), 4.86 (dd, J=11.6, 2.0 Hz, 1H), 4.48 (septet, J=6.7 Hz, 1H), 4.34 (br d, J=11.7 Hz, 1H), 3.92 (br d, J=11.7 Hz, 1H), 3.24-3.34 (br m, 1H), 3.06 (dd, J=12.9, 4.1 Hz, 1H), 2.65-2.73 (m, 1H), 2.32-2.46 (m, 1H), 1.96-2.06 (m, 1H), 1.49 and 1.49 (2 d, each J=6.6 Hz, each 3H).

Characteristic ¹H NMR peaks attributed to minor isomer C49: 6.20 (d, J=1.8 Hz, 1H), 4.68 (presumed septet, J=6.6 Hz, 1H), 1.46 (d, J=6.4 Hz, 3H), 1.45 (d, J=6.6 Hz, 3H).

Step 2. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-[1-(propan-2-yl)-1H-pyrazol-3-yl]-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (10)

The mixture of isomers from the preceding step (roughly 4:1 ratio, 35.8 mg, 72 μmol) was dissolved in methanol (5 mL) and treated with 1,8-diazabicyclo[5.4.0]undec-7-ene (8.6 μL, 58 μmol). The reaction mixture was heated at 55° C. for 4 hours, then allowed to stir at room temperature for 18 hours. After concentration in vacuo, purification was carried out via chromatography on silica gel (Gradient: 0% to 20% methanol in dichloromethane) followed by supercritical fluid chromatography [Column: Chiralpak IB, 5 μm; Eluent: 80:20 carbon dioxide/methanol containing 0.2% dimethylethylamine (v/v)] to afford the product. The indicated regiochemistry and relative stereochemistry of the pyrazole side chain for 10 were assigned via nuclear Overhauser enhancement studies. Yield: 1.9 mg, 4.8 μmol, 7%. LCMS m/z 393.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.41 (ddd, J=9.0, 9.0, 6.6 Hz, 1H), 7.35 (d, J=2.4 Hz, 1H), 6.87-6.94 (m, 1H), 6.82 (ddd, J=12.3, 8.4, 2.5 Hz, 1H), 6.29 (d, J=2.4 Hz, 1H), 4.79 (dd, J=11.6, 2.1 Hz, 1H), 4.49 (septet, J=6.7 Hz, 1H), 4.23 (dd, J=11.5, 2.0 Hz, 1H), 3.98 (d, J=11.7 Hz, 1H), 3.09-3.17 (m, 1H), 2.99 (dd, J=12.4, 4.0 Hz, 1H), 2.71 (dd, J=12.4, 2.6 Hz, 1H), 2.17-2.28 (m, 1H), 1.88 (ddd, J=13.3, 3.7, 2.3 Hz, 1H), 1.48 (d, J=6.6 Hz, 3H), 1.47 (d, J=6.6 Hz, 3H).

Example 11 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(2-methyl-1,3-oxazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-amine, hydrochloride salt (11)

Step 1. Synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carbonyl chloride (C50)

To a solution of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylic acid (C19) (580 mg, 1.34 mmol) in dichloromethane (6.7 mL) was added oxalyl chloride (0.253 mL, 2.95 mmol) drop-wise, followed by N,N-dimethylformamide (16 μL, 0.21 mmol). The reaction mixture was stirred for 30 minutes, then concentrated in vacuo. This material was used directly in the following step. Yield: 600 mg, 1.33 mmol, 99%. LCMS m/z 447.1 and 449.2 [M+H⁺ for corresponding methyl ester, due to reaction of acid chloride with methanol in the LCMS eluent]. ¹H NMR (400 MHz, CDCl₃) δ 8.39-8.43 (m, 2H), 7.66-7.71 (m, 1H), 7.50-7.62 (m, 3H), 7.04-7.10 (m, 1H), 6.95 (ddd, J=12.7, 8.0, 2.5 Hz, 1H), 4.51 (dd, J=11.8, 2.5 Hz, 1H), 4.46 (d, J=12.9 Hz, 1H), 4.18 (dd, J=12.8, 1.3 Hz, 1H), 3.36-3.44 (m, 1H), 3.09 (dd, J=13.6, 3.7 Hz, 1H), 2.85 (dd, J=13.7, 3.2 Hz, 1H), 2.36 (ddd, J=13.7, 4.5, 2.5 Hz, 1H), 2.10-2.21 (m, 1H).

Step 2. Synthesis of N-[(4aR,6R,8aS)-6-(bromoacetyl)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C51)

(4aR,6R,8aS)-2-(Benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carbonyl chloride (C50) (from the preceding step, 600 mg, 1.33 mmol) was dissolved in a 1:1 mixture of tetrahydrofuran and acetonitrile (6.7 mL) and added to a 0° C. solution of (diazomethyl)(trimethyl)silane (2 M in 1:1 tetrahydrofuran/acetonitrile, 2.33 mL, 4.66 mmol). After 2.5 hours, aqueous hydrobromic acid (48%, 1.51 mL, 13.3 mmol) was added drop-wise. The reaction mixture was stirred for 10 minutes, then quenched via addition of saturated aqueous sodium bicarbonate solution. The aqueous layer was extracted three times with ethyl acetate, and the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo. Chromatography on silica gel (Gradient: 0% to 40% ethyl acetate in heptane) afforded the product as a solid. Yield: 330 mg, 0.648 mmol, 49%. LCMS m/z 509.0, 511.0 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ 8.19 (br d, J=7 Hz, 2H), 7.48-7.54 (m, 1H), 7.34-7.47 (m, 3H), 6.85-6.97 (m, 2H), 4.26 (AB quartet, J_(AB)=14.1 Hz, Δν_(AB)=44.1 Hz, 2H), 3.11-3.19 (m, 1H), 3.00 (dd, J=13.1, 3.9 Hz, 1H), 2.68 (dd, J=13.0, 2.8 Hz, 1H), 2.04-2.17 (m, 2H).

Step 3. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(2-methyl-1,3-oxazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C52)

Acetamide (11.6 mg, 0.196 mmol) was added to a solution of N-[(4aR,6R,8aS)-6-(bromoacetyl)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C51) (25 mg, 49 μmol) in N,N-dimethylformamide (0.5 mL), and the reaction was heated to 110° C. for 1 hour, then cooled to room temperature and concentrated under reduced pressure. Purification via silica gel chromatography (Gradient: 0% to 100% ethyl acetate in heptane) provided the product as a solid. Yield: 8.1 mg, 17 μmol, 35%. LCMS m/z 470.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.21 (br d, J=7.6 Hz, 2H), 7.38-7.55 (m, 5H), 6.86-6.98 (m, 2H), 4.71 (br d, J=11 Hz, 1H), 4.31 (br d, J=12.3 Hz, 1H), 3.90 (d, J=12.1 Hz, 1H), 3.22-3.30 (m, 1H), 3.05 (dd, J=12.9, 4.1 Hz, 1H), 2.69 (dd, J=12.9, 2.9 Hz, 1H), 2.44 (s, 3H), 2.21-2.33 (m, 1H), 2.06-2.13 (m, 1H).

Step 4. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(2-methyl-1,3-oxazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine, hydrochloride salt (11)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(2-methyl-1,3-oxazol-4-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C52) was converted to the free base of the product according to the general procedure for the synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (1) in Example 1. Salt formation was carried out by dissolving the free base in dichloromethane, treating it with excess 1 M hydrogen chloride in diethyl ether, and removing solvents in vacuo. The product was obtained as a solid. Yield: 4.0 mg, 10 μmol, 53%. LCMS m/z 366.0 [M+H⁺]. ¹H NMR of the free base of 11: (400 MHz, CDCl₃) δ 7.53 (d, J=1.0 Hz, 1H), 7.42 (ddd, J=9.1, 8.9, 6.7 Hz, 1H), 6.77-6.89 (m, 2H), 4.66 (br dd, J=11.7, 2.0 Hz, 1H), 4.24 (dd, J=11.1, 2.3 Hz, 1H), 3.90 (d, J=11.2 Hz, 1H), 2.96-3.03 (m, 2H), 2.61-2.68 (m, 1H), 2.47 (s, 3H), 2.13-2.25 (m, 1H), 1.82 (ddd, J=13.5, 3.8, 2.5 Hz, 1H).

Example 12 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(4-methylpyrimidin-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (12)

Step 1. Synthesis of N-[(4aR,6R,8aS)-6-carbamimidoyl-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C53)

Trimethyloxonium tetrafluoroborate (98%, 48 mg, 0.32 mmol) was added to a suspension of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C43) (114 mg, 0.264 mmol) in dichloromethane (1.5 mL), and the reaction mixture was stirred at room temperature for 14 hours. During this time, additional trimethyloxonium tetrafluoroborate (27 mg, 0.18 mmol) was added. Solvents were removed in vacuo, and the residue was swirled with diethyl ether. The diethyl ether was decanted off, and the remaining material was transferred to a sealable tube with methanol (1 mL), treated with a solution of ammonia in methanol (7.0 M, 2 mL) and heated to 65° C. for 1 hour, then at 50° C. for 2 hours. The volume of the reaction mixture was reduced in vacuo, and ethyl acetate (100 mL) was added. The mixture was washed with saturated aqueous sodium bicarbonate solution (100 mL), and the aqueous layer was extracted with ethyl acetate (50 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (150 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure. Silica gel chromatography (Eluents: 95:4:1 ethyl acetate/methanol/concentrated ammonium hydroxide, followed by 80:18:2, then 75:25:2 ethyl acetate/methanol/concentrated ammonium hydroxide) provided the product as a white foam. By ¹H NMR, the product contained an impurity of related structure. Yield: 30 mg, 70 μmol, 26%. LCMS m/z 429.1 [M−H⁺]. ¹H NMR (400 MHz, CDCl₃), product peaks only: δ 8.14-8.19 (m, 2H), 7.50-7.56 (m, 1H), 7.31-7.48 (m, 3H), 6.85-6.97 (m, 2H), 4.49 (dd, J=11.7, 2.8 Hz, 1H), 4.19-4.25 (m, 1H), 3.87 (d, J=12.2 Hz, 1H), 3.16-3.25 (m, 1H), 2.96-3.02 (m, 1H), 2.66-2.73 (m, 1H), 2.08-2.29 (m, 2H).

Step 2. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(4-methylpyrimidin-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C54)

4,4-Dimethoxybutan-2-one (95%, 130 μL, 0.925 mmol) and a solution of sodium methoxide in methanol (1.0 M, 1.16 mL, 1.16 mmol) were added to a solution of N-[(4aR,6R,8aS)-6-carbamimidoyl-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C53) (99.9 mg, 0.232 mmol) in methanol (0.3 mL), and the reaction mixture was stirred at 50° C. for 2 hours. After it had cooled to room temperature, the reaction mixture was diluted with water and chloroform. The organic layer was washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (Gradient: 0% to 5% methanol in dichloromethane) afforded the product as a white solid. By ¹H NMR, the product contained an impurity of related structure. Yield: 34 mg, 71 μmol, 31%. LCMS m/z 481.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), product peaks only: δ 8.60 (d, J=5.1 Hz, 1H), 8.17-8.22 (m, 2H), 7.41-7.53 (m, 4H), 7.09 (br d, J=5.1 Hz, 1H), 6.87-6.98 (m, 2H), 4.93 (dd, J=11.6, 2.4 Hz, 1H), 4.40 (dd, J=12.2, 1.8 Hz, 1H), 4.06 (d, J=12.2 Hz, 1H), 3.3-3.38 (m, 1H), 3.07 (dd, J=13.0, 4.1 Hz, 1H), 2.71 (dd, J=12.9, 2.9 Hz, 1H), 2.55 (br s, 3H), 2.40-2.52 (m, 1H), 2.25 (ddd, J=13.8, 4.0, 2.5 Hz, 1H).

Step 3. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(4-methylpyrimidin-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (12)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(4-methylpyrimidin-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C54) was converted to the product according to the general procedure for the synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (1) in Example 1. The product was obtained as a white solid. Yield: 19 mg, 50 μmol, 81%. LCMS m/z 377.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.65 (d, J=5.2 Hz, 1H), 7.55 (ddd, J=9.1, 9.0, 6.7 Hz, 1H), 7.08 (br d, J=5.2 Hz, 1H), 6.85-6.91 (m, 1H), 6.81 (ddd, J=12.3, 8.6, 2.6 Hz, 1H), 4.90 (dd, J=11.6, 2.2 Hz, 1H), 4.31 (dd, J=11.0, 2.3 Hz, 1H), 4.08 (d, J=11.1 Hz, 1H), 3.06-3.14 (m, 1H), 3.01 (dd, J=12.3, 4.4 Hz, 1H), 2.64 (dd, J=12.2, 2.6 Hz, 1H), 2.58 (br s, 3H), 2.13-2.25 (m, 1H), 1.91 (ddd, J=13.2, 3.7, 2.5 Hz, 1H).

Example 13 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine, trifluoroacetate salt (13)

Step 1. Synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N—(N-hydroxyethanimidoyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C55)

The product was synthesized using the method described for synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C20) in Example 1, except that N′ hydroxyacetamidine was used in place of 2,2-dimethoxyethanamine. The product was obtained as a solid. Yield: 122 mg, 0.250 mmol, 53%. LCMS m/z 489.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.21 (br d, J=7 Hz, 2H), 7.50-7.57 (m, 1H), 7.37-7.49 (m, 3H), 6.87-6.99 (m, 2H), 4.45 (dd, J=12, 2.6 Hz, 1H), 4.18 (br d, J=12 Hz, 1H), 3.90 (br d, J=12 Hz, 1H), 3.16-3.24 (m, 1H), 3.03 (dd, J=13, 4 Hz, 1H), 2.67-2.75 (m, 1H), 2.26-2.38 (m, 1H), 2.10-2.2 (m, 1H), 1.96 (s, 3H).

Step 2. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C56)

A mixture of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(N-hydroxyethanimidoyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C55) (82 mg, 0.17 mmol), N,N-dimethylformamide (3 mL) and five 3-5 mm spherical beads of 3 Angstrom molecular sieves was heated in a microwave reactor at 140° C. for 1 hour. The solvent was removed using a Genevac system, and the crude product was combined with the product of an identical reaction carried out on C55 (40 mg, 82 μmol). Purification using silica gel chromatography (Gradient: 0% to 100% ethyl acetate in heptane) afforded the product as a white solid. Yield: 63 mg, 0.13 mmol, 52%. LCMS m/z 471.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) 8.16 (br d, J=7 Hz, 2H), 7.50-7.57 (m, 1H), 7.37-7.49 (m, 3H), 6.87-6.99 (m, 2H), 5.00 (dd, J=11.9, 2.5 Hz, 1H), 4.34 (dd, J=12.2, 1.4 Hz, 1H), 4.00 (d, J=11.9 Hz, 1H), 3.22-3.32 (m, 1H), 3.06 (dd, J=13.1, 3.9 Hz, 1H), 2.71 (dd, J=13.0, 2.7 Hz, 1H), 2.48-2.60 (m, 1H), 2.42 (s, 3H), 2.14-2.22 (m, 1H).

Step 3. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine, trifluoroacetate salt (13)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C56) (72.9 mg, 0.15 mmol) was combined with methanol (2.0 mL) and 1,8-diazabicyclo[5.4.0]undec-7-ene (26 μL, 0.17 mmol) and heated at reflux for 6 hours. The reaction mixture was cooled and concentrated in vacuo, then diluted with dichloromethane and washed with water. The aqueous layer was extracted twice with dichloromethane, and the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was purified via reversed-phase HPLC (Column: Waters Sunfire C18, 5 μm; Mobile phase A: 0.05% trifluoroacetic acid in water (v/v); Mobile phase B: 0.05% trifluoroacetic acid in acetonitrile (v/v); Gradient: 10% to 40% B) to provide the product. Yield: 27.9 mg, 76 μmol, 51%. LCMS m/z 367.1 [M+H⁺]. ¹H NMR (600 MHz, DMSO-d₆) δ 7.38-7.44 (m, 1H), 7.32-7.38 (m, 1H), 7.26 (ddd, J=8.3, 8.3, 2.2 Hz, 1H), 5.27 (dd, J=11.6, 2 Hz, 1H), 4.19 (d, J=12.3 Hz, 1H), 4.03 (d, J=12.3 Hz, 1H), 3.3-3.4 (m, 1H, assumed; obscured by water peak), 3.13 (dd, J=13.2, 2.6 Hz, 1H), 2.96 (dd, J=13.4, 4.2 Hz, 1H), 2.38 (s, 3H), 2.20-2.25 (m, 1H), 1.99-2.07 (m, 1H).

Example 14 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1,3,4-oxadiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (14)

Step 1. Synthesis of tert-butyl 2-{[(4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-6-yl]carbonyl}hydrazinecarboxylate (C57)

The product was synthesized using the method described for synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C20) in Example 1, except that tert-butyl hydrazinecarboxylate was used in place of 2,2-dimethoxyethanamine. The product was obtained as a white solid. Yield: 160 mg, 0.293 mmol, 83%. LCMS m/z 547.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ 7.50-7.58 (m, 1H), 7.34-7.50 (m, 3H), 6.86-7.00 (m, 2H), 4.33 (br d, J=12 Hz, 1H), 4.20 (d, J=12 Hz, 1H), 3.90 (d, J=12 Hz, 1H), 3.14-3.23 (m, 1H), 2.99-3.07 (m, 1H), 2.70 (d, J=12 Hz, 1H), 1.45 (s, 9H).

Step 2. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(hydrazinylcarbonyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C58)

Trifluoroacetic acid (0.6 mL) was added to a solution of tert-butyl 2-{[(4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-6-yl]carbonyl}hydrazinecarboxylate (C57) (160 mg, 0.293 mmol) in dichloromethane (0.9 mL), and the reaction mixture was stirred for 30 minutes. After concentration in vacuo, the residue was partitioned between dichloromethane and saturated aqueous sodium bicarbonate solution. The aqueous layer was extracted twice with dichloromethane, and the combined organic layers were dried over magnesium sulfate, filtered, and concentrated under reduced pressure to provide the product as a white solid. From ¹H NMR analysis, the product was characterized as a roughly 2:1 mixture of rotamers. Yield: 131 mg, 0.293 mmol, 100%. ¹H NMR (400 MHz, CDCl₃) δ 9.05 and 7.73 (2 br s, total 1H), 8.20 (br d, J=7 Hz, 2H), 7.49-7.55 (m, 1H), 7.34-7.47 (m, 3H), 6.85-6.98 (m, 2H), [4.33 (dd, J=11.9, 2.7 Hz) and 4.25 (dd, J=11.8, 2.6 Hz), total 1H], [4.22 (dd, J=12.3, 1.2 Hz) and 4.17 (dd, J=12.3, 1.4 Hz), total 1H], 3.84-3.95 (m, 1H), 3.12-3.22 (m, 1H), 2.98-3.04 (m, 1H), 2.66-2.74 (m, 1H), [2.33 (ddd, J=13.9, 4, 3 Hz) and 2.22 (ddd, J=13.9, 4, 3 Hz), total 1H], 2.02-2.17 (m, 1H).

Step 3. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,3,4-oxadiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C59)

p-Toluenesulfonic acid monohydrate (98%, 1.2 mg, 6.2 μmol) was added to a solution of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(hydrazinylcarbonyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-yl]benzamide (C58) (131 mg, 0.293 mmol) in trimethyl orthoformate (2 mL), and the reaction mixture was heated at 110° C. for 17 hours. After removal of volatiles in vacuo, chromatography on silica gel (Gradient: 0% to 100% ethyl acetate in heptane) afforded the product as a white solid. Yield: 36 mg, 79 μmol, 27%. LCMS m/z 457.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.43 (s, 1H), 8.17 (d, J=7.4 Hz, 2H), 7.51-7.56 (m, 1H), 7.37-7.49 (m, 3H), 6.88-6.99 (m, 2H), 5.09 (dd, J=11.9, 2.5 Hz, 1H), 4.35 (dd, J=12.2, 1.5 Hz, 1H), 3.97 (d, J=12.1 Hz, 1H), 3.24-3.32 (m, 1H), 3.07 (dd, J=13.1, 4.1 Hz, 1H), 2.72 (dd, J=13.0, 2.8 Hz, 1H), 2.57-2.69 (m, 1H), 2.12-2.19 (m, 1H).

Step 4. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1,3,4-oxadiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (14)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1,3,4-oxadiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C59) was converted to the product according to the method described for synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine, trifluoroacetate salt (13) in Example 13. The product was purified via reversed-phase HPLC (Column: Waters XBridge C18, 5 μm; Mobile phase A: 0.03% ammonium hydroxide in water (v/v); Mobile phase B: 0.03% ammonium hydroxide in acetonitrile (v/v); Gradient: 5% to 20% B). Yield: 15.9 mg, 45.1 μmol, 53%. LCMS m/z 353.0 [M+H⁺]. ¹H NMR (600 MHz, DMSO-d₆), characteristic peaks: δ 9.24 (s, 1H), 7.37 (ddd, J=9.2, 8.8, 7.0 Hz, 1H), 7.23 (ddd, J=12.5, 9.2, 2.4 Hz, 1H), 7.12 (ddd, J=8.8, 8.3, 2.6 Hz, 1H), 6.18 (br s, 2H), 5.03 (dd, J=11.8, 2.2 Hz, 1H), 4.11 (dd, J=10.7, 1.5 Hz, 1H), 3.67 (d, J=10.5 Hz, 1H), 2.86-2.92 (m, 1H), 2.19-2.28 (m, 1H), 1.91-1.96 (m, 1H).

Example 15 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(4,5-dimethyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-amine (15)

Step 1. Synthesis of 3-oxobutan-2-yl (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylate (C60)

1-[3-(Dimethylamino)propyl]-3-ethylcarbodimide hydrochloride (332 mg, 1.73 mmol) and 4-(dimethylamino)pyridine (14.2 mg, 0.116 mmol) were added to a solution of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylic acid (C19) (500 mg, 1.16 mmol) and 3-hydroxybutan-2-one (132 mg, 1.50 mmol) in dichloromethane (10 mL). After stirring for 3 days, the reaction mixture was diluted with additional dichloromethane and washed sequentially with saturated aqueous sodium bicarbonate solution and with water. The organic layer was dried over magnesium sulfate, filtered, and concentrated in vacuo to provide the crude product as a purple foam (815 mg), which was used directly in the following step. By ¹H NMR, this material consisted of a roughly 1:1 mixture of diastereomers at the methyl group adjacent to the ketone. LCMS m/z 503.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃), characteristic peaks: δ [5.21 (q, J=7.1 Hz) and 5.21 (q, J=7.1 Hz), total 1H], 2.19 and 2.19 (2 s, total 3H), [1.47 (d, J=7.0 Hz) and 1.46 (d, J=7.1 Hz), total 3H].

Step 2. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(4,5-dimethyl-1,3-oxazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (15)

3-Oxobutan-2-yl (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylate (C60) (material from the previous step, ≦1.16 mmol) and ammonium acetate (98%, 454 mg, 5.77 mmol) were combined in acetic acid (7 mL) and heated at reflux for 4 hours. The reaction mixture was allowed to cool to room temperature and was concentrated in vacuo; the residue was dissolved in dichloromethane and washed with saturated aqueous sodium carbonate solution and with water. The organic layer was dried over magnesium sulfate, filtered, and concentrated under reduced pressure. Silica gel chromatography (Gradient: 0% to 13% methanol in dichloromethane) afforded the product as a beige solid. Yield: 168 mg, 0.443 mmol, 38% over 2 steps. LCMS m/z 380.1 [M+H⁺]. ¹H NMR (400 MHz, CD₃OD) δ 7.33-7.41 (m, 1H), 6.93-7.03 (m, 2H), 4.73 (dd, J=11.9, 2.4 Hz, 1H), 4.24 (dd, J=11.2, 2.0 Hz, 1H), 3.77 (d, J=11.3 Hz, 1H), 2.98-3.05 (m, 1H), 2.91 (dd, half of ABX pattern, J=12.5, 4.1 Hz, 1H), 2.74 (dd, half of ABX pattern, J=12.6, 2.8 Hz, 1H), 2.31-2.42 (m, 1H), 2.24-2.26 (m, 3H), 2.06-2.08 (m, 3H), 1.83 (ddd, J=13.3, 3.9, 2.5 Hz, 1H).

Example 16 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(3-methyl-1,2-oxazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (16)

Step 1. Synthesis of methyl (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylate (C61)

To a stirring suspension of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylic acid (C19) (245 mg, 0.567 mmol) in dichloromethane (2.85 mL) was added oxalyl chloride (100 μL, 1.16 mmol) in a drop-wise fashion, followed by N,N-dimethylformamide (7.0 μL, 90 μmol). The reaction mixture was stirred for 15 minutes, at which time additional oxalyl chloride (50 μL, 0.58 mmol) was added. After 20 minutes, methanol (1 mL) was added, and the reaction mixture was stirred for 10 minutes. Removal of solvents in vacuo was followed by silica gel chromatography (Gradient: 0% to 60% ethyl acetate in heptane), affording the product as a white solid. Yield: 208 mg, 0.466 mmol, 82%. LCMS m/z 447.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.18 (br d, J=7 Hz, 2H), 7.50-7.56 (m, 1H), 7.36-7.49 (m, 3H), 6.85-6.97 (m, 2H), 4.33 (dd, J=12.0, 2.6 Hz, 1H), 4.19 (dd, J=12.2, 1.6 Hz, 1H), 3.94 (d, J=12.3 Hz, 1H), 3.80 (s, 3H), 3.13-3.22 (m, 1H), 3.03 (dd, half of ABX pattern, J=13.1, 4.0 Hz, 1H), 2.67 (dd, half of ABX pattern, J=13.0, 2.8 Hz, 1H), 2.21-2.34 (m, 1H), 2.05-2.12 (m, 1H).

Step 2. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(3-methyl-1,2-oxazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C62)

To a solution of propan-2-one oxime (23.0 mg, 0.315 mmol) in tetrahydrofuran (1.25 mL) at 0° C. was added a solution of n-butyllithium in hexanes (2.5 M, 0.25 mL, 0.62 mmol). The ice bath was removed, and the mixture was stirred at room temperature for 30 minutes. After the reaction mixture had been re-cooled to 0° C., a solution of methyl (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxylate (C61) (70.0 mg, 0.157 mmol) in tetrahydrofuran (0.75 mL) was added drop-wise. The reaction mixture was allowed to warm to room temperature and stirred for 1 hour, then cooled once again in an ice bath. Concentrated sulfuric acid (35 μL, 0.66 mmol) was added, and the flask was allowed to warm to room temperature for 1 hour. After re-cooling the reaction mixture to 0° C., it was neutralized with 5 M aqueous sodium hydroxide solution. Water (2 mL) was added, and the mixture was extracted with ethyl acetate (3×5 mL). The combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (Gradient: 0% to 50% ethyl acetate in heptane) provided the product as a white solid; the relative stereochemistry of the isoxazole side chain was confirmed via nuclear Overhauser enhancement study. Yield: 18 mg, 38 μmol, 24%. LCMS m/z 470.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.18-8.22 (m, 2H), 7.50-7.55 (m, 1H), 7.38-7.48 (m, 3H), 6.88-6.99 (m, 2H), 6.14 (s, 1H), 4.88 (dd, J=11.7, 2.4 Hz, 1H), 4.31 (dd, J=12.1, 1.6 Hz, 1H), 3.93 (d, J=12.3 Hz, 1H), 3.24-3.31 (m, 1H), 3.06 (dd, J=12.9, 4.1 Hz, 1H), 2.70 (dd, J=13.0, 2.8 Hz, 1H), 2.28 (s, 3H), 2.26-2.38 (m, 1H), 2.11 (ddd, J=13.6, 4.2, 2.7 Hz, 1H).

Step 3. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(3-methyl-1,2-oxazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (16)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(3-methyl-1,2-oxazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C62) was converted to the product according to the method described for synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(1-methyl-1H-pyrazol-3-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-amine (2) in Example 2. The product was obtained as a white solid. Yield: 8.9 mg, 24 μmol, 80%. LCMS m/z 366.1 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 7.36 (ddd, J=9.0, 9.0, 6.6 Hz, 1H), 6.86-6.92 (m, 2H), 6.83 (ddd, J=12.5, 8.6, 2.5 Hz, 1H), 4.82 (dd, J=11.9, 2.4 Hz, 1H), 4.23 (dd, J=11.2, 2.2 Hz, 1H), 3.91 (d, J=11.2 Hz, 1H), 2.98-3.08 (m, 2H), 2.65-2.71 (m, 1H), 2.30 (s, 3H), 2.11-2.22 (m, 1H), 1.91-1.98 (m, 1H).

Example 17 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(5-methyl-1,3,4-oxadiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-amine (17)

Step 1. Synthesis of N-[(4aR,6R,8aS)-6-[(2-acetylhydrazinyl)carbonyl]-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C63)

The product was synthesized using the method described for synthesis of (4aR,6R,8aS)-2-(benzoylamino)-8a-(2,4-difluorophenyl)-N-(2,2-dimethoxyethyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazine-6-carboxamide (C20) in Example 1, except that acetohydrazide was used in place of 2,2-dimethoxyethanamine. The product was obtained as a white solid. Yield: 128 mg, 0.262 mmol, 95%. LCMS m/z 489.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.7-8.9 (v br s, 1H), 8.19 (br d, J=7 Hz, 2H), 7.85-8.1 (v br s, 1H), 7.50-7.57 (m, 1H), 7.42-7.49 (m, 2H), 7.38 (ddd, J=9, 9, 6 Hz, 1H), 6.86-6.98 (m, 2H), 4.34 (dd, J=11.8, 3.0 Hz, 1H), 4.20 (br d, J=12.2 Hz, 1H), 3.91 (d, J=12.2 Hz, 1H), 3.12-3.22 (br m, 1H), 3.02 (dd, J=13.0, 3.9 Hz, 1H), 2.68 (dd, J=13.0, 2.5 Hz, 1H), 2.08-2.26 (m, 2H), 2.06 (s, 3H).

Step 2. Synthesis of N-[(4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(5-methyl-1,3,4-oxadiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C64)

Trifluoromethanesulfonic anhydride (99%, 0.129 mL, 0.759 mmol) was added drop-wise over 1 to 2 minutes to a 0° C. solution of triphenylphosphine oxide (99%, 106 mg, 0.378 mmol) in dichloromethane (2 mL), and the solution was stirred for 5 minutes. The ice bath was then removed, and the reaction mixture was allowed to warm to room temperature. A solution of N-[(4aR,6R,8aS)-6-[(2-acetylhydrazinyl)carbonyl]-8a-(2,4-difluorophenyl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C63) (123 mg, 0.252 mmol) in dichloromethane (1.5 mL) was added, and stirring was continued for 30 minutes. After the reaction was quenched with 10% aqueous sodium bicarbonate solution, the aqueous layer was extracted twice with dichloromethane. The combined organic layers were dried over magnesium sulfate, filtered, and concentrated in vacuo. Purification via silica gel chromatography (Gradient: 0% to 5% methanol in dichloromethane) afforded the product as a colorless oil. Yield: 79 mg, 0.17 mmol, 67%. LCMS m/z 471.2 [M+H⁺]. ¹H NMR (400 MHz, CDCl₃) δ 8.17 (br d, J=7.3 Hz, 2H), 7.50-7.55 (m, 1H), 7.36-7.48 (m, 3H), 6.87-6.98 (m, 2H), 5.00 (dd, J=11.8, 2.4 Hz, 1H), 4.32 (dd, J=12.2, 1.5 Hz, 1H), 3.95 (d, J=12.2 Hz, 1H), 3.22-3.30 (m, 1H), 3.05 (dd, J=13.0, 4.0 Hz, 1H), 2.71 (dd, J=13.1, 2.8 Hz, 1H), 2.53-2.65 (m, 1H), 2.53 (s, 3H), 2.10 (ddd, J=13.8, 4.0, 2.5 Hz, 1H).

Step 3. Synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(5-methyl-1,3,4-oxadiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-amine (17)

N-[(4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(5-methyl-1,3,4-oxadiazol-2-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-][1,3]thiazin-2-yl]benzamide (C64) was converted to the product according to the method described for synthesis of (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(3-methyl-1,2,4-oxadiazol-5-yl)-4,4-a,5,6,8,8a-hexahydropyrano[3,4-d][1,3]thiazin-2-amine, trifluoroacetate salt (13) in Example 13. The product was subjected to reversed-phase HPLC (Column: Waters XBridge C18, 5 μm; Mobile phase A: 0.03% ammonium hydroxide in water (v/v); Mobile phase B: 0.03% ammonium hydroxide in acetonitrile (v/v); Gradient: 20% to 50% B). Yield: 30.5 mg, 83.2 μmol, 49%. LCMS m/z 367.0 [M+H⁺]. ¹H NMR (600 MHz, DMSO-d₆) δ 7.36 (ddd, J=9.2, 8.8, 7.4 Hz, 1H), 7.22 (ddd, J=12.7, 9, 2.4 Hz, 1H), 7.11 (ddd, J=8.8, 8.3, 2.6 Hz, 1H), 6.16 (br s, 2H), 4.93 (dd, J=11.8, 2.2 Hz, 1H), 4.09 (dd, J=10.7, 1.5 Hz, 1H), 3.65 (d, J=10.5 Hz, 1H), 2.85-2.90 (m, 1H), 2.76 (d, J=3.5 Hz, 2H), 2.51 (s, 3H), 2.17-2.25 (m, 1H), 1.86-1.91 (m, 1H).

TABLE 6 Method of preparation; ¹H NMR (400 MHz, CDCl₃), δ (ppm); Example starting Mass spectrum, observed ion m/z Number material(s) Structure (M + 1) (unless otherwise indicated) 18 Ex 9; C25¹

¹H NMR (400 MHz, CD₃CN) δ 9.09 (d, J = 1.4 Hz, 1H), 8.73 (d, J = 5.2 Hz, 1H), 7.52 (ddd, J = 5.2, 1.5, 0.7 Hz, 1H), 7.34-7.42 (m, 1H), 6.95-7.03 (m, 2H), 4.72 (dd, J = 11.7, 2.5 Hz, 1H), 4.23 (dd, J = 11.2, 2.0 Hz, 1H), 3.88 (d, J = 11.2 Hz, 1H), 3.07-3.14 (m, 1H). 2.91 (dd, J = 12.7, 4.0 Hz, 1H), 2.74 (dd, J = 12.8, 3.0 Hz, 1H), 2.10 (ddd, J = 13.3, 4.0, 2.6 Hz, 1H), 1.77-1.88 (m, 1H); 363.1 19 Ex 10; C25²

7.41 (ddd, J = 9.0, 9.0, 6.5 Hz, 1H), 7.37 (d, J = 2.4 Hz, 1H), 6.86-6.93 (m, 1H), 6.82 (ddd, J = 12.3, 8.4, 2.5 Hz, 1H), 6.27 (d, J = 2.4 Hz, 1H), 4.76 (dd, J = 11.5, 2.2 Hz, 1H), 4.22 (dd, J = 11.5, 2.2 Hz, 1H), 3.97 (d, J = 11.5 Hz, 1H), 3.53-3.59 (m, 1H), 3.06-3.14 (m, 1H), 2.99 (dd, J = 12.5, 4.1 Hz, 1H), 2.70 (dd, J = 12.5, 2.7 Hz, 1H), 2.16-2.27 (m, 1H), 1.88 (ddd, J = 13.5, 3.9, 2.4 Hz, 1H), 0.97-1.04 (m, 2H), 1.04-1.12 (m, 2H); 391.2 20 Ex 4; C19

¹H NMR (500 MHz, CD₃OD) δ 8.50 (q, J = 1.5 Hz, 1H), 7.35-7.41 (m, 1H), 6.96-7.04 (m, 2H), 4.93 (dd, J = 11.9, 2.6 Hz, 1H), 4.28 (dd, J = 11.2, 1.8 Hz, 1H), 3.81 (d, J = 11.2, 1H), 3.06-3.12 (m, 1H), 2.94 (dd, half of ABX pattern, J = 12.6, 4.2 Hz, 1H), 2.78 (dd, half of ABX pattern, J = 12.6, 2.9 Hz, 1H), 2.34-2.43 (m, 1H), 1.96 (ddd, J = 13.4, 3.9, 2.8 Hz, 1H); 420.1 21 Ex 12; C53³

8.80 (d, J = 4.9 Hz, 2H), 7.56 (ddd, J = 9.1, 9.0, 6.8 Hz, 1H), 7.24 (t, J = 4.9 Hz, 1H), 6.85-6.91 (m, 1H), 6.81 (ddd, J = 12.4, 8.6, 2.6 Hz, 1H), 4.94 (dd, J = 11.7, 2.4 Hz, 1H), 4.32 (dd, J = 11.0, 2.2 Hz, 1H), 4.06 (d, J = 11.0 Hz, 1H), 3.06-3.14 (m, 1H), 3.03 (dd, J = 12.3, 4.3 Hz, 1H), 2.64 (dd, J = 12.3, 2.6 Hz, 1H), 2.14-2.25 (m, 1H), 1.90 (ddd, J = 13.1, 3.7, 2.5 Hz, 1H); 363.2 22 Ex 4; C33

¹H NMR (400 MHz, CD₃OD) δ 7.60 (q, J = 1.3 Hz, 1H), 7.32-7.39 (m, 1H), 6.95-7.04 (m, 2H), 4.79 (dd, J = 11.7, 2.8 Hz, 1H), 4.28 (dd, J = 11.2, 2.0 Hz, 1H), 3.82 (d, J = 11.2 Hz, 1H), 3.11- 3.18 (m, 1H), 2.89 (ddd, J = 12.0, 4, 4 Hz, 1H), 2.15 (d, J = 1.3 Hz, 3H), 1.98- 2.10 (m, 1H), 1.94 (ddd, half of ABXY pattern, J = 13.3, 4.2, 3.0 Hz, 1H), 1.21 (d, J = 6.9 Hz, 3H); 380.1 23 Ex 12; C53⁴

8.65 (d, J = 0.4 Hz, 2H), 7.50 (ddd, J = 9.1, 9.0, 6.7 Hz, 1H), 6.87-6.93 (m, 1H), 6.83 (ddd, J = 12.4, 8.6, 2.6 Hz, 1H), 4.96 (dd, J = 11.7, 2.4 Hz, 1H), 4.31 (dd, J = 11.2, 2.2 Hz, 1H), 4.07 (d, J = 11.4 Hz, 1H), 3.09-3.17 (m, 1H), 3.04 (dd, J = 12.3, 4.2 Hz, 1H), 2.68 (dd, J = 12.4, 2.7 Hz, 1H), 2.16-2.27 (m, 1H), 1.92-1.98 (m, 1H); 381.2 24 Ex 12; C53⁵

¹H NMR (600 MHz, DMSO-d₆) δ 8.69 (s, 2H), 7.34-7.43 (m, 2H), 7.23-7.28 (m, 1H), 4.89 (dd, J = 11.6, 2.4 Hz, 1H), 4.07 (AB quartet, J_(AB) = 12.3 Hz, Δν_(AB) = 71 Hz, 2H), 3.3-3.4 (m, 1H, assumed; obscured by water peak), 3.10 (dd, J = 13.2, 2.6 Hz, 1H), 2.97 (dd, J = 12.9, 3.7 Hz, 1H), 2.29 (s, 3H), 2.10-2.18 (m, 1H), 2.00-2.05 (m, 1H); 377.0 ¹The relative stereochemistry of the pyrimidine side chain was confirmed via nuclear Overhauser enhancement study. ²The indicated structure for Example 19 was assigned via nuclear Overhauser enhancement studies. ³In this case, 3-(dimethylamino)prop-2-enal was used in place of 4,4-dimethoxybutan-2-one. ⁴3-(Dimethylamino)-2-fluoroprop-2-enal (see K. England et al., Tetrahedron Lett. 2010, 51, 2849-2851) was used in place of 4,4-dimethoxybutan-2-one. ⁵3-(Dimethylamino)-2-methylprop-2-enal was used in place of 4,4-dimethoxybutan-2-one.

Biological Assays BACE1 Cell-Free Assay

Beta-secretase (BACE) is one of the enzymes involved in the generation of the amyloid beta peptide found in the amyloid plaques of Alzheimer's Disease patients. This assay measures the inhibition of the beta-secretase enzyme as it cleaves a non-native peptide.

A synthetic APP substrate that can be cleaved by beta-secretase having N-terminal biotin and made fluorescent by the covalent attachment of Oregon Green at the Cys residue is used to assay beta-secretase activity in the presence or absence of the inhibitory compounds. The substrate is Biotin-GLTNIKTEEISEISY^EVEFR-C[Oregon Green]KK-OH. The BACE1 enzyme is affinity purified material from conditioned media of CHO-K1 cells that have been transfected with a soluble BACE construct (BACE1deltaTM96His). Compounds are incubated in a ½ log dose response curve from a top concentration of 100 μM with BACE1 enzyme and the biotinylated fluorescent peptide in 384-well black plates (Thermo Scientific #4318). BACE1 is at a final concentration of 0.1 nM with a final concentration of peptide substrate of 150 nM in a reaction volume of 30 μL assay buffer (100 mM sodium acetate, pH 4.5 (brought to pH with acetic acid), and 0.001% Tween-20). Plates are covered and incubated for 3 hours at 37° C. The reaction is stopped with the addition of 30 μL of 1.5 μM Streptavidin (Pierce, #21125). After a 10 minute incubation at room temperature, plates are read on a PerkinElmer Envision for fluorescent polarization (Ex485 nm/Em530 nm). The activity of the beta-secretase enzyme is detected by changes in the fluorescence polarization that occur when the substrate is cleaved by the enzyme. Incubation in the presence of compound inhibitor demonstrates specific inhibition of beta-secretase enzymatic cleavage of the synthetic APP substrate.

Whole Cell-Free Assay (In Vitro sAPPb Assay):

H4 human neuroglioma cells over-expressing the wild-type human APP₆₉₅ are treated for 18 hours with compound in a final concentration 1% DMSO. sAPPβ levels are measured using TMB-ELISA with capture APP N-terminal antibody (Affinity BioReagents, OMA1-03132), wild-type sAPPβ specific reporter p192 (Elan), and tertiary anti rabbit-HRP (GE Healthcare).

BACE2 Assay:

This assay measures the inhibition of the BACE2 enzyme as it cleaves a non-native peptide. A synthetic substrate that can be cleaved by BACE2 having N-terminal biotin and made fluorescent by the covalent attachment of Oregon Green at the Cys residue is used to assay BACE2 activity in the presence or absence of the inhibitory compounds. The substrate is Biotin-KEISEISYEVEFR-C(Oregon green)-KK-OH. The BACE2 enzyme is available from Enzo Life Sciences (Cat # BML-SE550). Compounds are incubated in a ½ log dose response curve from a top concentration of 100 μM with BACE2 enzyme and the biotinylated fluorescent peptide in 384-well black plates (Thermo Scientific #4318). BACE2 is at a final concentration of 2.5 nM with a final concentration of peptide substrate of 150 nM in a reaction volume of 30 μL assay buffer (100 mM Sodium Acetate, pH 4.5 (brought to pH with acetic acid), and 0.001% Tween-20). Plates are covered and incubated for 3 hours at 37° C. The reaction is stopped with the addition of 30 μL of 1.5 μM Streptavidin (Pierce, #21125). After a 10 minute incubation at room temperature, plates are read on a PerkinElmer Envision for fluorescent polarization (Ex485 nm/Em530 nm). The activity of the beta-secretase enzyme is detected by changes in the fluorescence polarization that occur when the substrate is cleaved by the enzyme. Incubation in the presence of compound inhibitor demonstrates specific inhibition of BACE2 enzymatic cleavage of the synthetic substrate.

TABLE 7 BACE1 sAPPβ BACE2 Cell-free Whole-Cell Cell-free Example Assay IC₅₀ Assay IC₅₀ Assay number IUPAC Name (μM)^(a) (nM)^(a) IC₅₀ (μM)^(b) 1 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6- 0.511 32 0.619 (1,3-oxazol-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 2 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1- 0.183 17 4.74 methyl-1H-pyrazol-3-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 3 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1- 7.78 636   N.D.^(c) methyl-1H-pyrazol-5-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 4 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(4- 0.084  4 0.273 methyl-1,3-oxazol-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 5 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(5- 0.473 23 1.15 methyl-1,3-oxazol-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 6 (4R,4aR,6R,8aS)-8a-(2,4-Difluorophenyl)- 0.198 32 2.37 4-methyl-6-(1,3-oxazol-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 7 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(1- 0.055  5 0.185 methyl-1H-pyrazol-4-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 8 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(4- 0.260 27 9.72 methyl-1,3-thiazol-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 9 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(2- 0.583 28 >100 methylpyrimidin-4-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 10 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-[1- N.D. N.D. N.D. (propan-2-yl)-1H-pyrazol-3-yl]- 4,4a,5,6,8,8a-hexahydropyrano[3,4- d][1,3]thiazin-2-amine 11 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(2- 0.316 34 2.02 methyl-1,3-oxazol-4-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine, hydrochloride salt 12 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(4- 1.47 50 N.D. methylpyrimidin-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 13 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(3- 0.439 34 0.208 methyl-1,2,4-oxadiazol-5-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine, trifluoroacetate salt 14 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6- 2.21 196  N.D. (1,3,4-oxadiazol-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 15 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6- 0.112  7 2.82 (4,5-dimethyl-1,3-oxazol-2-yl)- 4,4a,5,6,8,8a-hexahydropyrano[3,4- d][1,3]thiazin-2-amine 16 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(3- 0.059  3 4.55 methyl-1,2-oxazol-5-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 17 (4aR,6R,8aS)-8a-(2,4-Difluorophenyl)-6-(5- 0.301 10 5.56 methyl-1,3,4-oxadiazol-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 18 (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6- 0.390 28 0.695 (pyrimidin-4-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 19 (4aR,6R,8aS)-6-(1-cyclopropyl-1H-pyrazol- 0.444 N.D. N.D. 3-yl)-8a-(2,4-difluorophenyl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 20 (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-[4- 0.467 14 0.629 (trifluoromethyl)-1,3-oxazol-2-yl]- 4,4a,5,6,8,8a-hexahydropyrano[3,4- d][1,3]thiazin-2-amine 21 (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6- 0.983 44 3.09 (pyrimidin-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 22 (4R,4aR,6R,8aS)-8a-(2,4-difluorophenyl)- 0.089  6 1.40 4-methyl-6-(4-methyl-1,3-oxazol-2-yl)- 4,4a,5,6,8,8a-hexahydropyrano[3,4- d][1,3]thiazin-2-amine 23 (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(5- 0.161^(b)  10^(b) N.D. fluoropyrimidin-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine 24 (4aR,6R,8aS)-8a-(2,4-difluorophenyl)-6-(5- 0.688 24 1.08 methylpyrimidin-2-yl)-4,4a,5,6,8,8a- hexahydropyrano[3,4-d][1,3]thiazin-2- amine, trifluoroacetate salt ^(a)Reported IC₅₀ values are the geometric mean of 2-7 determinations. ^(b)IC₅₀ value is from a single determination. ^(c)Not determined 

We claim:
 1. A compound having the absolute stereochemistry of Formula I

wherein R¹ is hydrogen or methyl, wherein said methyl is optionally substituted with one to three fluoro; R² is a 5- to 6-membered heteroaryl, having one to three heteroatoms selected from N, O or S, wherein at least one of the heteroatoms is S or N and wherein said N is optionally substituted with R⁴; and wherein said heteroaryl moiety is independently substituted at each available carbon position with R^(3a), R^(3b), R^(3c) or R^(3d); R^(3a), R^(3b), R^(3c) and R^(3d) are independently hydrogen, halogen, —OH, —CN, C₁₋₆alkyl, —OC₁₋₆alkyl; —(CR^(5a)R^(5b))_(m)—C₃₋₆cycloalkyl or —(CR^(5a)R^(5b))_(m)—C₃₋₆heterocycloalkyl; wherein said alkyl is optionally substituted with one to three fluoro and wherein said cycloalkyl and heterocycloalkyl moieties are optionally substituted with one to three fluoro, —CH₃, —CH₂F, —CHF₂ or —CF₃; R⁴ is hydrogen, C₁₋₆alkyl, or —(CR^(5a)R^(5b))_(m)—C₃₋₆cycloalkyl; wherein said alkyl and cycloalkyl are optionally substituted with one to three fluoro, —OCH₃ or —OCF₃; R^(5a) and R^(5b) are independently hydrogen, —CH₃, —CH₂F, —CHF₂, —CF₃ or —OCH₃; and m is 0, 1 or 2; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 2. The compound of claim 1 wherein at least one of the heteroatoms of the R² group is N; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 3. The compound of claim 2 wherein R² is selected from the group consisting of

or a tautomer thereof or pharmaceutically acceptable salt of said compound or tautomer.
 4. The compound of claim 3 wherein R^(3a), R^(3b), R^(3c) and R^(3d) are independently hydrogen, fluoro, C₁₋₆alkyl or —(CR^(5a)R^(5b))_(m)—C₃₋₆cycloalkyl, wherein said alkyl and cycloalkyl are optionally substituted with one to three fluoro; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 5. The compound of claim 4 wherein m is 0; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 6. The compound of claim 2 wherein R² is selected from the group consisting of

and wherein R^(3a), R^(3b) or R^(3c) are independently hydrogen, halogen, C₁₋₆alkyl or —(CR^(5a)R^(5b))_(m)—C₃₋₆cycloalkyl wherein said alkyl or cycloalkyl are optionally substituted with one to three fluoro; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 7. The compound of claim 6 wherein R⁴ is hydrogen, methyl, isopropyl or cyclopropyl and m is 0; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 8. The compound of claim 2 wherein R² is

R^(3a) and R^(3b) are independently hydrogen, halogen, C₁₋₆alkyl or —(CR^(5a)R^(5b))_(m)—C₃₋₆cycloalkyl, wherein said alkyl and cycloalkyl are optionally substituted with one to three fluoro; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 9. The compound of claim 8 wherein R^(3a) is independently hydrogen or methyl and R^(3b) is independently hydrogen or methyl; wherein said methyl moieties are optionally substituted with one to three fluoro; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 10. The compound of claim 8 wherein the compound is selected from

or a tautomer thereof or a pharmaceutically acceptable salt of said tautomer or compound.
 11. The compound of claim 2 wherein R² is selected from

R^(3a) and R^(3b) are independently hydrogen, halogen, C₁₋₆alkyl or —(CR^(5a)R^(5b))_(m)—C₃₋₆cycloalkyl, wherein said alkyl and cycloalkyl are optionally substituted with one to three fluoro; R⁴ is hydrogen, C₁₋₆alkyl, or —(CR^(5a)R^(5b))_(m)—C₃₋₆cycloalkyl; wherein said alkyl and cycloalkyl are optionally substituted with one to three fluoro; and wherein m is zero; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 12. The compound of claim 11 wherein R^(3a) and R^(3b) are hydrogen or C₁₋₆alkyl; and R⁴ is hydrogen, methyl, isopropyl or cyclopropyl; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 13. The compound of claim 12 selected from

or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 14. The compound of claim 2 wherein R² is selected from

R^(3a), R^(3b) and R^(3c) are independently hydrogen, halogen, C₁₋₆alkyl or C₃₋₆cycloalkyl, wherein said alkyl and cycloalkyl are optionally substituted with one to three fluoro; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 15. The compound of claim 14 selected from

or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 16. The compound of claim 2 wherein R² is selected from

wherein R^(3a) and R^(3b) are independently hydrogen, halogen, C₁₋₆alkyl or C₃₋₆cycloalkyl, wherein said alkyl and cycloalkyl are optionally substituted with one to three fluoro; or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 17. The compound of claim 16 selected from

or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer.
 18. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 1, or a tautomer thereof or a pharmaceutically acceptable salt of said compound or tautomer, and a pharmaceutically acceptable vehicle, diluent or carrier. 